TOHOKU UNIVERSITY Researchers

Details of the Researcher

Home

日本語 English

Keiko Nakayama

Section

Graduate School of Medicine

Job title

Professor

Degree

医学博士（東京医科歯科大学）

researchmap

https://researchmap.jp/Tohoku_KEIKON

J-GLOBAL ID

200901043258496930

Research History 11

2018/10 - Present

Tohoku University
2003/01 - Present

東北大学大学院医学系研究科　教授
2012/04 - 2015/03

東北大学メディカル・メガバンク機構　人材育成部門長
1997/05 - 2003/01

九州大学生体防御医学研究所　助教授
1997/05 - 2002/12

九州大学生体防御医学研究所　助教授
1996/08 - 1997/04

東京医科歯科大学医学部付属病院　医院
1996/06 - 1997/04

東京医科歯科大学　医員
1995/07 - 1996/07

日本ロシュ株式会社　研究員
1995/07 - 1996/05

日本ロシュ　主任研究員
1991/04 - 1995/06

米国ワシントン大学　ポストドクタルフェロー
1991/04 - 1995/06

ワシントン大学医学部内科アレルギー免疫部門　博士研究員

Show all Show first 5

Education 2

Tokyo Medical and Dental University　Graduate School, Division of Medicine　Internal Medicine

- 1991/03
Tokyo Medical and Dental University　Faculty of Medicine　Medical School

- 1986/03

Committee Memberships 2

復興促進プログラム（A－STEP）評価委員会　委員

2014/04 - 2015/03
復興促進プログラム（A－STEP）評価委員会　委員

2014/04 - 2015/03

Professional Memberships 5

日本癌学会(2007/04-2007/09 プログラム委員)
日本細胞生物学会(2007/05-2010/04 プログラム委員)
日本細胞生物学会(2006/11-2007/05 将来検討委員)
日本分子生物学会(1996/04-)
日本癌学会(1999/04-)

Research Interests 3

genetic engineering
protein degradation
cell cycle

Research Areas 3

Life sciences / Developmental biology /
Life sciences / Cell biology /
Life sciences / Molecular biology /

Awards 3

FIRSTシンポジウム「『科学技術が拓く2030年』へのシナリオ」内NEXTライフ・イノベーション・ポスターセッション銅賞

2013/02/28　RIRSTプログラム公開活動実行委員会
平成１９年度医学部・医学系研究科「教育貢献賞」

2008/03　東北大学医学部長、医学系研究科長
東京医科歯科大学研究奨励賞

1992/04　東京医科歯科大学

Papers 283

Biallelic variants/mutations of IL1RAP in patients with steroid-sensitive nephrotic syndrome. International-journal Peer-reviewed

Sou Niitsuma, Hiroki Kudo, Atsuo Kikuchi, Takaya Hayashi, Satoshi Kumakura, Shuhei Kobayashi, Yuko Okuyama, Naonori Kumagai, Tetsuya Niihori, Yoko Aoki, Takanori So, Ryo Funayama, Keiko Nakayama, Matsuyuki Shirota, Shuji Kondo, Shoji Kagami, Hiroyasu Tsukaguchi, Kazumoto Iijima, Shigeo Kure, Naoto Ishii

International immunology　32　(4)　283-292　2019/12/24

DOI： 10.1093/intimm/dxz081 　

More details Close

Nephrotic syndrome (NS) is a renal disease characterized by severe proteinuria and hypoproteinemia. Although several single-gene mutations have been associated with steroid-resistant NS, causative genes for steroid-sensitive NS (SSNS) have not been clarified. While seeking to identify causative genes associated with SSNS by whole-exome sequencing, we found compound heterozygous variants/mutations (c.524T>C; p.I175T and c.662G>A; p.R221H) of the interleukin-1 receptor accessory protein (IL1RAP) gene in two siblings with SSNS. The siblings' parents are healthy, and each parent carries a different heterozygous IL1RAP variant/mutation. Since IL1RAP is a critical subunit of the functional interleukin-1 receptor (IL-1R), we investigated the effect of these variants on IL-1R subunit function. When stimulated with IL-1β, peripheral blood mononuclear cells from the siblings with SSNS produced markedly lower levels of cytokines compared with cells from healthy family members. Moreover, IL-1R with a variant IL1RAP subunit, reconstituted on a hematopoietic cell line, had impaired binding ability and low reactivity to IL-1β. Thus, the amino acid substitutions in IL1RAP found in these NS patients are dysfunctional variants/mutations. Furthermore, in the kidney of Il1rap-/- mice, the number of myeloid-derived suppressor cells, which require IL-1β for their differentiation, was markedly reduced although these mice did not show significantly increased proteinuria in acute nephrotic injury with lipopolysaccharide treatment. Together, these results identify two IL1RAP variants/mutations in humans for the first time and suggest that IL1RAP might be a causative gene for familial NS.
Heme regulates protein interactions and phosphorylation of BACH2 intrinsically disordered region in humoral response

Miki Watanabe-Matsui, Shun Kadoya, Kei Segawa, Hiroki Shima, Tadashi Nakagawa, Yuko Nagasawa, Shuichiro Hayashi, Mitsuyo Matsumoto, Mariko Ikeda, Akihiko Muto, Kyoko Ochiai, Long C. Nguyen, Katsumi Doh-Ura, Mikako Shirouzu, Keiko Nakayama, Kazutaka Murayama, Kazuhiko Igarashi

iScience　28　(1)　111529-111529　2025/01
Publisher: Elsevier BV
DOI： 10.1016/j.isci.2024.111529 　

ISSN： 2589-0042
Updated Genetic Analysis of Japanese Familial ALS Patients Carrying SOD1 Variants Revealed Phenotypic Differences for Common Variants. International-journal

Ayumi Nishiyama, Tetsuya Niihori, Naoki Suzuki, Rumiko Izumi, Tetsuya Akiyama, Masaaki Kato, Ryo Funayama, Keiko Nakayama, Hitoshi Warita, Yoko Aoki, Masashi Aoki

Neurology. Genetics　10　(6)　e200196　2024/12

DOI： 10.1212/NXG.0000000000200196 　

More details Close

BACKGROUND AND OBJECTIVES: Amyotrophic lateral sclerosis (ALS) is an adult-onset progressive neurodegenerative disease. Approximately 10% of ALS cases are familial, and more than 20 causative genes have been identified. As we have previously reported, SOD1 variants are the most common causes of familial ALS in Japan. Because antisense oligonucleotides for SOD1-linked ALS are being used in practical applications, the types of variants and the clinical features of patients need to be updated. METHODS: We consecutively recruited 160 families with familial ALS in Japan. We performed genetic analyses, focusing on SOD1-linked ALS as the most common in our cohort, updated their genotypes, and characterized clinical phenotypes. RESULTS: A total of 26 SOD1 variants in 56 patients and 49 families (30.6%) were collected, with the 3 most common (p.His47Arg [the conventional numbering; H46R], p.Leu127Ser [L126S], p.Asn87Ser [N86S]) accounting for 38.8% of all families. We also identified 2 novel variants (p.Ile36Phe [I35F] and p.Asn132Argfs*3 [N131Rfs*3]). The mean age at onset was 48.9 ± 12.2 (mean ± SD) years for all patients with SOD1-linked ALS. Lower limb onset comprised 70% of cases. The mean disease duration was 64.7 ± 82 months, and the median survival was 71.5 months. Some variants led to a relatively homogeneous phenotype, although clinical characteristics differed among types of variants and families. Patients with p.His47Arg (H46R) showed slower progression with lower limb onset and a predominance of lower motor neuron involvement. The p.Leu127Ser (L126S) variant led to varying degrees of progression in heterozygous or homozygous states and presented incomplete penetrance. Intrafamilial phenotypic differences were observed in families carrying p.Asn87Ser (N86S). Four variants (p.Cys7Gly [C6G], p.His44Arg [H43R], p.Leu85Val [L84V], and p.Cys147Arg [C146R]) were found to be associated with rapid disease progression. DISCUSSION: The genetic basis of familial ALS, at least for SOD1 variants, still differed by geographic and ethnic background. Understanding these clinical profiles will help optimize evaluation in targeted gene therapy worldwide and benefit efficient diagnosis, leading to precise application in clinical practice.
Calreticulin exposure induced by anticancer drugs is associated with the p53 signaling pathway in colorectal cancer cells

Satoru Naito, Taiki Kajiwara, Hideaki Karasawa, Tomoyuki Ono, Tatsushi Saito, Ryo Funayama, Keiko Nakayama, Shinobu Ohnuma, Michiaki Unno

Biochemical and Biophysical Research Communications　733　150665-150665　2024/11
Publisher: Elsevier BV
DOI： 10.1016/j.bbrc.2024.150665 　

ISSN： 0006-291X
ALS-associated RNA binding proteins converge onUNC13Atranscription through regulation of REST

Yasuaki Watanabe, Naoki Suzuki, Tadashi Nakagawa, Masaki Hosogane, Tetsuya Akiyama, Hitoshi Warita, Masashi Aoki, Keiko Nakayama

2024/10/23

DOI： 10.1101/2024.10.22.619761 　
Accelerated plasma-cell differentiation in Bach2-deficient mouse B cells is caused by altered IRF4 functions. International-journal

Kyoko Ochiai, Hiroki Shima, Toru Tamahara, Nao Sugie, Ryo Funayama, Keiko Nakayama, Tomohiro Kurosaki, Kazuhiko Igarashi

The EMBO journal　2024/04/11

DOI： 10.1038/s44318-024-00077-6 　

More details Close

Transcription factors BACH2 and IRF4 are both essential for antibody class-switch recombination (CSR) in activated B lymphocytes, while they oppositely regulate the differentiation of plasma cells (PCs). Here, we investigated how BACH2 and IRF4 interact during CSR and plasma-cell differentiation. We found that BACH2 organizes heterochromatin formation of target gene loci in mouse splenic B cells, including targets of IRF4 activation such as Aicda, an inducer of CSR, and Prdm1, a master plasma-cell regulator. Release of these gene loci from heterochromatin in response to B-cell receptor stimulation was coupled to AKT-mTOR pathway activation. In Bach2-deficient B cells, PC genes' activation depended on IRF4 protein accumulation, without an increase in Irf4 mRNA. Mechanistically, a PU.1-IRF4 heterodimer in activated B cells promoted BACH2 function by inducing gene expression of Bach2 and Pten, a negative regulator of AKT signaling. Elevated AKT activity in Bach2-deficient B cells resulted in IRF4 protein accumulation. Thus, BACH2 and IRF4 mutually modulate the activity of each other, and BACH2 inhibits PC differentiation by both the repression of PC genes and the restriction of IRF4 protein accumulation.
TANK Binding Kinase 1 Promotes BACH1 Degradation through Both Phosphorylation-Dependent and -Independent Mechanisms without Relying on Heme and FBXO22. International-journal

Liang Liu, Mitsuyo Matsumoto, Miki Watanabe-Matsui, Tadashi Nakagawa, Yuko Nagasawa, Jingyao Pang, Bert K K Callens, Akihiko Muto, Kyoko Ochiai, Hirotaka Takekawa, Mahabub Alam, Hironari Nishizawa, Mikako Shirouzu, Hiroki Shima, Keiko Nakayama, Kazuhiko Igarashi

International journal of molecular sciences　25　(8)　2024/04/09

DOI： 10.3390/ijms25084141 　

More details Close

BTB and CNC homology 1 (BACH1) represses the expression of genes involved in the metabolism of iron, heme and reactive oxygen species. While BACH1 is rapidly degraded when it is bound to heme, it remains unclear how BACH1 degradation is regulated under other conditions. We found that FBXO22, a ubiquitin ligase previously reported to promote BACH1 degradation, polyubiquitinated BACH1 only in the presence of heme in a highly purified reconstitution assay. In parallel to this regulatory mechanism, TANK binding kinase 1 (TBK1), a protein kinase that activates innate immune response and regulates iron metabolism via ferritinophagy, was found to promote BACH1 degradation when overexpressed in 293T cells. While TBK1 phosphorylated BACH1 at multiple serine and threonine residues, BACH1 degradation was observed with not only the wild-type TBK1 but also catalytically impaired TBK1. The BACH1 degradation in response to catalytically impaired TBK1 was not dependent on FBXO22 but involved both autophagy-lysosome and ubiquitin-proteasome pathways judging from its suppression by using inhibitors of lysosome and proteasome. Chemical inhibition of TBK1 in hepatoma Hepa1 cells showed that TBK1 was not required for the heme-induced BACH1 degradation. Its inhibition in Namalwa B lymphoma cells increased endogenous BACH1 protein. These results suggest that TBK1 promotes BACH1 degradation in parallel to the FBXO22- and heme-dependent pathway, placing BACH1 as a downstream effector of TBK1 in iron metabolism or innate immune response.
Downregulation of ABCC3 activates MAPK signaling through accumulation of deoxycholic acid in colorectal cancer cells. International-journal

Yukihiro Sato, Minoru Kobayashi, Masahiro Ohira, Ryo Funayama, Masamitsu Maekawa, Hideaki Karasawa, Ryosuke Kashiwagi, Yayoi Aoyama, Nariyasu Mano, Shinobu Ohnuma, Michiaki Unno, Keiko Nakayama

Cancer science　2024/04/02

DOI： 10.1111/cas.16132 　

More details Close

ABCC3 (also known as MRP3) is an ATP binding cassette transporter for bile acids, whose expression is downregulated in colorectal cancer through the Wnt/β-catenin signaling pathway. However, it remained unclear how downregulation of ABCC3 expression contributes to colorectal carcinogenesis. We explored the role of ABCC3 in the progression of colorectal cancer-in particular, focusing on the regulation of bile acid export. Gene expression analysis of colorectal adenoma isolated from familial adenomatous polyposis patients revealed that genes related to bile acid secretion including ABCC3 were downregulated as early as at the stage of adenoma formation. Knockdown or overexpression of ABCC3 increased or decreased intracellular concentration of deoxycholic acid, a secondary bile acid, respectively, in colorectal cancer cells. Forced expression of ABCC3 suppressed deoxycholic acid-induced activation of MAPK signaling. Finally, we found that nonsteroidal anti-inflammatory drugs increased ABCC3 expression in colorectal cancer cells, suggesting that ABCC3 could be one of the targets for therapeutic intervention of familial adenomatous polyposis. Our data thus suggest that downregulation of ABCC3 expression contributes to colorectal carcinogenesis through the regulation of intracellular accumulation of bile acids and activity of MAPK signaling.
Nuclear pore pathology underlying multisystem proteinopathy type 3-related inclusion body myopathy. International-journal

Rumiko Izumi, Kensuke Ikeda, Tetsuya Niihori, Naoki Suzuki, Matsuyuki Shirota, Ryo Funayama, Keiko Nakayama, Hitoshi Warita, Maki Tateyama, Yoko Aoki, Masashi Aoki

Annals of clinical and translational neurology　2023/12/29

DOI： 10.1002/acn3.51977 　

More details Close

OBJECTIVE: Multisystem proteinopathy type 3 (MSP3) is an inherited, pleiotropic degenerative disorder caused by a mutation in heterogeneous nuclear ribonucleoprotein A1 (hnRNPA1), which can affect the muscle, bone, and/or nervous system. This study aimed to determine detailed histopathological features and transcriptomic profile of HNRNPA1-mutated skeletal muscles to reveal the core pathomechanism of hereditary inclusion body myopathy (hIBM), a predominant phenotype of MSP3. METHODS: Histopathological analyses and RNA sequencing of HNRNPA1-mutated skeletal muscles harboring a c.940G > A (p.D314N) mutation (NM_031157) were performed, and the results were compared with those of HNRNPA1-unlinked hIBM and control muscle tissues. RESULTS: RNA sequencing revealed aberrant alternative splicing events that predominantly occurred in myofibril components and mitochondrial respiratory complex. Enrichment analyses identified the nuclear pore complex (NPC) and nucleocytoplasmic transport as suppressed pathways. These two pathways were linked by the hub genes NUP50, NUP98, NUP153, NUP205, and RanBP2. In immunohistochemistry, these nucleoporin proteins (NUPs) were mislocalized to the cytoplasm and aggregated mostly with TAR DNA-binding protein 43 kDa and, to a lesser extent, with hnRNPA1. Based on ultrastructural observation, irregularly shaped myonuclei with deep invaginations were frequently observed in atrophic fibers, consistent with the disorganization of NPCs. Additionally, regarding the expression profiles of overall NUPs, reduced expression of NUP98, NUP153, and RanBP2 was shared with HNRNPA1-unlinked hIBMs. INTERPRETATION: The shared subset of altered NUPs in amyotrophic lateral sclerosis (ALS), as demonstrated in prior research, HNRNPA1-mutated, and HNRNPA1-unlinked hIBM muscle tissues may provide evidence regarding the underlying common nuclear pore pathology of hIBM, ALS, and MSP.
Neonatal developmental and epileptic encephalopathy with movement disorders and arthrogryposis: A case report with a novel missense variant of SCN1A. International-journal

Yukimune Okubo, Moriei Shibuya, Haruhiko Nakamura, Aritomo Kawashima, Kaori Kodama, Wakaba Endo, Takehiko Inui, Noriko Togashi, Yu Aihara, Matsuyuki Shirota, Ryo Funayama, Tetsuya Niihori, Atsushi Fujita, Keiko Nakayama, Yoko Aoki, Naomichi Matsumoto, Shigeo Kure, Atsuo Kikuchi, Kazuhiro Haginoya

Brain & development　2023/07/11

DOI： 10.1016/j.braindev.2023.06.009 　

More details Close

Variants of SCN1A represent the archetypal channelopathy associated with several epilepsy syndromes. The clinical phenotypes have recently expanded from Dravet syndrome. CASE REPORT: We present a female patient with the de novo SCN1A missense variant, c.5340G > A (p. Met1780Ile). The patient had various clinical features with neonatal onset SCN1A epileptic encephalopathy, arthrogryposis multiplex congenita, thoracic hypoplasia, thoracic scoliosis, and hyperekplexia. CONCLUSION: Our findings are compatible with neonatal developmental and epileptic encephalopathy with movement disorders and arthrogryposis; the most severe phenotype probably caused by gain-of-function variant of SCN1A. The efficacy of sodium channel blocker was also discussed. Further exploration of the phenotype-genotype relationship of SCN1A variants may lead to better pharmacological treatments and family guidance.
Genomic adaptive potential to cold environments in the invasive red swamp crayfish

Daiki X. Sato, Yuki Matsuda, Nisikawa Usio, Ryo Funayama, Keiko Nakayama, Takashi Makino

iScience　107267-107267　2023/07
Publisher: Elsevier BV
DOI： 10.1016/j.isci.2023.107267 　

ISSN： 2589-0042
Sex-Specific Differences in the Transcriptome of the Human Dorsolateral Prefrontal Cortex in Schizophrenia. International-journal

Zhiqian Yu, Kazuko Ueno, Ryo Funayama, Mai Sakai, Naoki Nariai, Kaname Kojima, Yoshie Kikuchi, Xue Li, Chiaki Ono, Junpei Kanatani, Jiro Ono, Kazuya Iwamoto, Kenji Hashimoto, Kengo Kinoshita, Keiko Nakayama, Masao Nagasaki, Hiroaki Tomita

Molecular neurobiology　60　(2)　1083-1098　2022/11/22

DOI： 10.1007/s12035-022-03109-6 　

More details Close

Schizophrenia presents clinical and biological differences between males and females. This study investigated transcriptional profiles in the dorsolateral prefrontal cortex (DLPFC) using postmortem data from the largest RNA-sequencing (RNA-seq) database on schizophrenic cases and controls. Data for 154 male and 113 female controls and 160 male and 93 female schizophrenic cases were obtained from the CommonMind Consortium. In the RNA-seq database, the principal component analysis showed that sex effects were small in schizophrenia. After we analyzed the impact of sex-specific differences on gene expression, the female group showed more significantly changed genes compared with the male group. Based on the gene ontology analysis, the female sex-specific genes that changed were overrepresented in the mitochondrion, ATP (phosphocreatine and adenosine triphosphate)-, and metal ion-binding relevant biological processes. An ingenuity pathway analysis revealed that the differentially expressed genes related to schizophrenia in the female group were involved in midbrain dopaminergic and γ-aminobutyric acid (GABA)-ergic neurons and microglia. We used methylated DNA-binding domain-sequencing analyses and microarray to investigate the DNA methylation that potentially impacts the sex differences in gene transcription using a maternal immune activation (MIA) murine model. Among the sex-specific positional genes related to schizophrenia in the PFC of female offspring from MIA, the changes in the methylation and transcriptional expression of loci ACSBG1 were validated in the females with schizophrenia in independent postmortem samples by real-time PCR and pyrosequencing. Our results reveal potential genetic risks in the DLPFC for the sex-dependent prevalence and symptomology of schizophrenia.
A novel variant in the transmembrane 4 domain of ANO3 identified in a two-year-old girl with developmental delay and tremor. International-journal

Yu Aihara, Matsuyuki Shirota, Atsuo Kikuchi, Yu Katata, Yu Abe, Tetsuya Niihori, Ryo Funayama, Keiko Nakayama, Yoko Aoki, Shigeo Kure

Journal of human genetics　2022/09/27

DOI： 10.1038/s10038-022-01082-5 　

More details Close

ANO3 encodes Anoctamin-3, also known as TMEM16C, a calcium-activated chloride channel. Heterozygous variants of ANO3 can cause dystonia 24, an adult-onset focal dystonia. Some pediatric cases have been reported, but most patients were intellectually normal with some exceptions. Here, we report a two-year-old girl who showed mild to moderate developmental delay, tremor, and ataxic gait, but no obvious dystonia. Trio exome sequencing identified a heterozygous de novo missense variant NM_031418.4:c.1809T>G, p.(Asn603Lys) in the ANO3 gene. Three cases with ANO3 variants and intellectual disability have been reported, including the present case. These variants were predicted to face in the same direction on the same alpha-helix (the transmembrane 4 domain), suggesting an association between these variants and childhood-onset movement disorder with intellectual disability. In pediatric cases with developmental delay and movement disorders such as tremor and ataxia, specific variants in the transmembrane 4 domain of ANO3 may be a cause, even in the absence of dystonia.
High levels of chromosomal instability facilitate the tumor growth and sphere formation. International-journal

Kenji Iemura, Hayato Anzawa, Ryo Funayama, Runa Iwakami, Keiko Nakayama, Kengo Kinoshita, Kozo Tanaka

Cancer science　2022/06/05

DOI： 10.1111/cas.15457 　

More details Close

Most cancer cells show chromosomal instability (CIN), a condition in which chromosome missegregation occurs at high rates. Growing evidence suggests that CIN is not just a consequence, but a driving force for oncogenic transformation, although the relationship between CIN and tumorigenesis has not been fully elucidated. Here we found that conventional two-dimensional (2D) culture of HeLa cells, a cervical cancer-derived cell line, is a heterogenous population containing cells with different CIN levels. Although cells with high CIN levels (high-CIN cells) grew slower than cells with low CIN levels (low-CIN cells) in 2D monolayer culture, they formed tumors in nude mice and larger spheres in three-dimensional (3D) culture, which is more representative of the in vivo environment. The duration of mitosis was longer in high-CIN cells, implicating their higher mitotic defects. Single cell genome sequencing revealed that high-CIN cells exhibit a higher karyotype heterogeneity compared with low-CIN cells. Intriguingly, the karyotype heterogeneity was reduced in the spheres formed by high-CIN cells, suggesting that cells with growth advantages were selected, although genomic copy number changes specific for spheres were not identified. When we examined gene expression profiles, genes related to K-ras signaling were upregulated, while those related to unfolded protein response were downregulated in high-CIN cells in 3D culture compared with 2D culture, suggesting the relevance of these genes for their survival. Our data suggest that although CIN is disadvantageous in monolayer culture, it promotes the selection of cells with growth advantages under in vivo environments, which may lead to tumorigenesis.
Novel POLE mutations identified in patients with IMAGE-I syndrome cause aberrant subcellular localisation and protein degradation in the nucleus. International-journal

Tomohiro Nakano, Yoji Sasahara, Atsuo Kikuchi, Kunihiko Moriya, Hidetaka Niizuma, Tetsuya Niihori, Matsuyuki Shirota, Ryo Funayama, Keiko Nakayama, Yoko Aoki, Shigeo Kure

Journal of medical genetics　59　(11)　1116-22　2022/05/09

DOI： 10.1136/jmedgenet-2021-108300 　

More details Close

BACKGROUND: DNA replisome is a molecular complex that plays indispensable roles in normal DNA replication. IMAGE-I syndrome is a DNA replisome-associated genetic disease caused by biallelic mutations in the gene encoding DNA polymerase epsilon catalytic subunit 1 (POLE). However, the underlying molecular mechanisms remain largely unresolved. METHODS: The clinical manifestations in two patients with IMAGE-I syndrome were characterised. Whole-exome sequencing was performed and altered mRNA splicing and protein levels of POLE were determined. Subcellular localisation, cell cycle analysis and DNA replication stress were assessed using fibroblasts and peripheral blood from the patients and transfected cell lines to determine the functional significance of POLE mutations. RESULTS: Both patients presented with growth retardation, adrenal insufficiency, immunodeficiency and complicated diffuse large B-cell lymphoma. We identified three novel POLE mutations: namely, a deep intronic mutation, c.1226+234G>A, common in both patients, and missense (c.2593T>G) and in-frame deletion (c.711_713del) mutations in each patient. The unique deep intronic mutation produced aberrantly spliced mRNAs. All mutants showed significantly reduced, but not null, protein levels. Notably, the mutants showed severely diminished nuclear localisation, which was rescued by proteasome inhibitor treatment. Functional analysis revealed impairment of cell cycle progression and increase in the expression of phospho-H2A histone family member X in both patients. CONCLUSION: These findings provide new insights regarding the mechanism via which POLE mutants are highly susceptible to proteasome-dependent degradation in the nucleus, resulting in impaired DNA replication and cell cycle progression, a characteristic of DNA replisome-associated diseases.
SPT16 ubiquitylation by DCAF14-CRL4 regulates FACT binding to histones. International-journal

Tadashi Nakagawa, Akane Morohoshi, Yuko Nagasawa, Makiko Nakagawa, Masaki Hosogane, Yasuhiro Noda, Toru Hosoi, Keiko Nakayama

Cell reports　38　(12)　110541-110541　2022/03/22

DOI： 10.1016/j.celrep.2022.110541 　

More details Close

The histone chaperone complex FACT comprises SPT16 and SSRP1 and contributes to DNA replication, transcription, and repair, but how it plays such various roles is unclear. Here, we show that human SPT16 is ubiquitylated at lysine-674 (K674) by the DCAF14-CRL4 ubiquitin ligase. K674 is located in the middle domain of SPT16, and the corresponding residue of the yeast ortholog is critical for binding to histone H3.1-H4. We show that the middle domain of human SPT16 binds to histone H3.1-H4 and that this binding is inhibited by K674 ubiquitylation. Cells with heterozygous knockin of a K674R mutant of SPT16 manifest reduction of both SPT16 ubiquitylation and H3.1 in chromatin, a reduced population in mid S phase, impaired proliferation, and increased susceptibility to S phase stress. Our data thus indicate that SPT16 ubiquitylation by DCAF14-CRL4 regulates FACT binding to histones and may thereby control DNA replication-coupled histone incorporation into chromatin.
幅広い臨床症状を呈したMECOM遺伝子変異を同定した2家系

新堀哲也, 田野島玲大, 笹原洋二, 佐藤篤, 入江正寛, 南條由佳, 舟山亮, 城田松之, 阿部太紀, 奥山祐子, 石井直人, 中山啓子, 呉繁夫, 今泉益栄, 青木洋子

日本小児科学会雑誌　126　(2)　230-230　2022/02
Publisher: (公社)日本小児科学会
ISSN： 0001-6543
Heterozygous calcyclin-binding protein/Siah1-interacting protein (CACYBP/SIP) gene pathogenic variant linked to a dominant family with paucity of interlobular bile duct. International-journal

Miyako Kanno, Mitsuyoshi Suzuki, Ken Tanikawa, Chikahiko Numakura, Shu-Ichi Matsuzawa, Tetsuya Niihori, Yoko Aoki, Yoichi Matsubara, Satoshi Makino, Gen Tamiya, Satoshi Nakano, Ryo Funayama, Matsuyuki Shirota, Keiko Nakayama, Tetsuo Mitsui, Kiyoshi Hayasaka

Journal of human genetics　67　(7)　393-397　2022/01/28

DOI： 10.1038/s10038-022-01017-0 　

More details Close

Paucity of interlobular bile ducts (PILBD) is a heterogeneous disorder classified into two categories, syndromic and non-syndromic bile duct paucity. Syndromic PILBD is characterized by the presence of clinical manifestations of Alagille syndrome. Non-syndromic PILBD is caused by multiple diseases, such as metabolic and genetic disorders, infectious diseases, and inflammatory and immune disorders. We evaluated a family with a dominantly inherited PILBD, who presented with cholestasis at 1-2 months of age but spontaneously improved by 1 year of age. Next-generation sequencing analysis revealed a heterozygous CACYBP/SIP p.E177Q pathogenic variant. Calcyclin-binding protein and Siah1 interacting protein (CACYBP/SIP) form a ubiquitin ligase complex and induce proteasomal degradation of non-phosphorylated β-catenin. Immunohistochemical analysis revealed a slight decrease in CACYBP and β-catenin levels in the liver of patients in early infancy, which almost normalized by 13 months of age. The CACYBP/SIP p.E177Q pathogenic variant may form a more active or stable ubiquitin ligase complex that enhances the degradation of β-catenin and delays the maturation of intrahepatic bile ducts. Our findings indicate that accurate regulation of the β-catenin concentration is essential for the development of intrahepatic bile ducts and CACYBP/SIP pathogenic variant is a novel cause of PILDB.
Phenotypic heterogeneity in individuals with MECOM variants in 2 families. International-journal

Tetsuya Niihori, Reo Tanoshima, Yoji Sasahara, Atsushi Sato, Masahiro Irie, Yuka Saito-Nanjo, Ryo Funayama, Matsuyuki Shirota, Taiki Abe, Yuko Okuyama, Naoto Ishii, Keiko Nakayama, Shigeo Kure, Masue Imaizumi, Yoko Aoki

Blood advances　6　(18)　5257-5261　2022/01/12

DOI： 10.1182/bloodadvances.2020003812 　

More details Close

MECOM encodes the transcriptional regulators, EVI1 and MDS1-EVI1, from two distinct transcription start sites. EVI1 plays important roles in hematopoiesis and stem cell self-renewal. Recently, our group and others revealed that individuals with MECOM variants present diverse hematological and skeletal defects, including radioulnar synostosis (RUS). In the present study, we analyzed two families suspected with MECOM-associated syndrome. In family 1, a MECOM splicing variant (c.2285+1G>A) was identified in an individual with bone marrow failure (TRS4) without RUS and her mother, who had mild leukocytopenia, thrombocytopenia, and bilateral RUS. A copy neutral loss of heterozygosity decreasing the variant allele frequency was observed in the bone marrow of TRS4 and the peripheral blood leukocytes of her mother. However, TRS4 remained transfusion-dependent. In family 2, a MECOM variant (c.2208-4A>G), which was predicted to cause a cryptic acceptor site that results in a 3-base insertion (an insertion of Ser) in the mRNA, was identified in the proband, with bone marrow failure; this variant was also observed in her brother and father, both of whom have skeletal malformations, but no cytopenia. RT-PCR using leukocytes revealed a transcript with a 3-bp insertion in the proband, her brother, and the father, suggesting that the transcript variant with a 3-bp insertion is independent of blood phenotype. Collectively, these results suggest the presence of intrafamilial clinical heterogeneity in both families with MECOM splicing variants. Somatic genetic event may complicate the understanding of clinical variability among family members.
Neurobehavioral characteristics of mice with SETD5 mutations as models of IDD23 and KBG syndromes. International-journal

Tadashi Nakagawa, Satoko Hattori, Toru Hosoi, Keiko Nakayama

Frontiers in genetics　13　1022339-1022339　2022

DOI： 10.3389/fgene.2022.1022339 　

More details Close

Genomic analysis has revealed that the genes for various chromatin regulators are mutated in many individuals with neurodevelopmental disorders (NDDs), emphasizing the important role of chromatin regulation in nervous system development and function. Chromatin regulation is mediated by writers, readers, and erasers of histone and DNA modifications, with such proteins being defined by specific domains. One of these domains is the SET domain, which is present in enzymes that catalyze histone methylation. Heterozygous loss-of-function mutations of the SETD5 (SET domain containing 5) gene have been identified in individuals with an NDD designated IDD23 (intellectual developmental disorder, autosomal dominant 23). KBG syndrome (named after the initials of the last names of the first three families identified with the condition) is characterized by features that either overlap with or are distinct from those of IDD23 and was initially thought to be caused only by mutations in the ANKRD11 (ankyrin repeat domain containing 11) gene. However, recent studies have identified SETD5 mutations in some KBG syndrome patients without ANKRD11 mutations. Here we summarize the neurobehavioral characterization of Setd5 +/- mice performed by four independent research groups, compare IDD23 and KBG phenotypes, and address the utility and future development of mouse models for elucidation of the mechanisms underlying NDD pathogenesis, with a focus on SETD5 and its related proteins.
Wnt5a in cancer-associated fibroblasts promotes colorectal cancer progression. International-journal

Tomoaki Hirashima, Hideaki Karasawa, Takashi Aizawa, Takashi Suzuki, Akihiro Yamamura, Hideyuki Suzuki, Taiki Kajiwara, Hiroaki Musha, Ryo Funayama, Matsuyuki Shirota, Shinobu Ohnuma, Keiko Nakayama, Michiaki Unno

Biochemical and biophysical research communications　568　37-42　2021/09/03

DOI： 10.1016/j.bbrc.2021.06.062 　

More details Close

Cancer-associated fibroblasts (CAFs) are a major component of the tumor microenvironment and have been shown to promote cancer aggressiveness. In our previous study, analysis of expression profiles obtained from paired CAFs and normal fibroblasts from colorectal cancer (CRC) tissue revealed that gene sets related to the Wnt signaling pathway were highly enriched in colorectal CAFs. Furthermore, among the components of the β-catenin-independent Wnt pathway, Wnt5a was highly expressed in CAFs. Since Wnt5a is considered to be a regulator of CRC progression in CAFs, we performed immunohistochemical analysis on Wnt5a in 171 patients who underwent surgery for CRC. Positive staining for Wnt5a was often found in cancer stroma, particularly in fibromatous areas, although the immunoreactivity for Wnt5a was weak in cancer cells. Wnt5a status in CAFs was significantly associated with tumor size, depth of invasion, lymphatic and vascular invasion, lymph node metastasis, TNM stage, and recurrence. Subsequent in vitro analyses using human recombinant Wnt5a protein revealed that cancer cell proliferation and migration were significantly increased by stimulation with Wnt5a. Our findings suggest that Wnt5a-derived CAFs play a crucial role in CRC progression and have potential as a target of anti-cancer therapies.
LSS欠損症における組織特異的モデルマウスを用いた代謝および病理プロファイル

和田陽一, 菊池敦生, 加賀元宗, 清水直紀, 伊藤隼哉, 新堀哲也, 佐藤孝太, 中澤徹, 中山啓子, 青木洋子, 仲川清隆, 呉繁夫

日本先天代謝異常学会雑誌　37　126-126　2021/09
Publisher: (一社)日本先天代謝異常学会
ISSN： 0912-0122
幹細胞から赤血球への通り道メチオニン代謝によるエピゲノム調節をかいした赤血球造血制御(Methionine metabolism controls erythropoiesis by epigenetic regulation)

加藤浩貴, Long Nugyen, 石井悠翔, 松本光代, 三枝大輔, 舟山亮, 岡江寛明, 藤原亨, 武藤哲彦, 中山啓子, 有馬隆博, Scadden David, 五十嵐和彦, 張替秀郎

日本血液学会学術集会　83回　PSY-4　2021/09
Publisher: (一社)日本血液学会
LSS欠損症における組織特異的モデルマウスを用いた代謝および病理プロファイル

和田陽一, 菊池敦生, 加賀元宗, 清水直紀, 伊藤隼哉, 新堀哲也, 佐藤孝太, 中澤徹, 中山啓子, 青木洋子, 仲川清隆, 呉繁夫

日本先天代謝異常学会雑誌　37　126-126　2021/09
Publisher: (一社)日本先天代謝異常学会
ISSN： 0912-0122
Alternative microexon splicing by RBFOX2 and PTBP1 is associated with metastasis in colorectal cancer. International-journal

Yasushi Mochizuki, Ryo Funayama, Matsuyuki Shirota, Yuna Kikukawa, Masahiro Ohira, Hideaki Karasawa, Minoru Kobayashi, Shinobu Ohnuma, Michiaki Unno, Keiko Nakayama

International journal of cancer　149　(10)　1787-1800　2021/08/04

DOI： 10.1002/ijc.33758 　

More details Close

The splicing of microexons (very small exons) is frequently dysregulated in the brain of individuals with autism spectrum disorder. However, little is known of the patterns, regulatory mechanisms, and roles of microexon splicing in cancer. We here examined the transcriptome-wide profile of microexon splicing in matched colorectal cancer (CRC) and normal tissue specimens. Out of 1492 microexons comprising 3 to 15 nucleotides, 21 (1%) manifested differential splicing between CRC and normal tissue. The 21 genes harboring the differentially spliced microexons were enriched in gene ontology terms related to cell adhesion and migration. RNA interference-mediated knockdown experiments identified two splicing factors, RBFOX2 and PTBP1, as regulators of microexon splicing in CRC cells. RBFOX2 and PTBP1 were found to directly bind to microexon-containing pre-mRNAs and to control their splicing in such cells. Differential microexon splicing was shown to be due, at least in part, to altered expression of RBFOX2 and PTBP1 in CRC tissue compared with matched normal tissue. Finally, we found that changes in the pattern of microexon splicing were associated with CRC metastasis. Our data thus suggest that altered expression of RBFOX2 and PTBP1 might influence CRC metastasis through the regulation of microexon splicing.
Reduced PHOX2B stability causes axonal growth impairment in motor neurons with TARDBP mutations. International-journal

Shio Mitsuzawa, Naoki Suzuki, Tetsuya Akiyama, Mitsuru Ishikawa, Takefumi Sone, Jiro Kawada, Ryo Funayama, Matsuyuki Shirota, Hiroaki Mitsuhashi, Satoru Morimoto, Kensuke Ikeda, Tomomi Shijo, Akiyuki Ohno, Naoko Nakamura, Hiroya Ono, Risako Ono, Shion Osana, Tadashi Nakagawa, Ayumi Nishiyama, Rumiko Izumi, Shohei Kaneda, Yoshiho Ikeuchi, Keiko Nakayama, Teruo Fujii, Hitoshi Warita, Hideyuki Okano, Masashi Aoki

Stem cell reports　16　(6)　1527-1541　2021/06/08

DOI： 10.1016/j.stemcr.2021.04.021 　

More details Close

Amyotrophic lateral sclerosis (ALS) is an adult-onset incurable motor neuron (MN) disease. The reasons for selective MN vulnerability in ALS are unknown. Axonal pathology is among the earliest signs of ALS. We searched for novel modulatory genes in human MN axon shortening affected by TARDBP mutations. In transcriptome analysis of RNA present in the axon compartment of human-derived induced pluripotent stem cell (iPSC)-derived MNs, PHOX2B (paired-like homeobox protein 2B) showed lower expression in TARDBP mutant axons, which was consistent with axon qPCR and in situ hybridization. PHOX2B mRNA stability was reduced in TARDBP mutant MNs. Furthermore, PHOX2B knockdown reduced neurite length in human MNs. Finally, phox2b knockdown in zebrafish induced short spinal axons and impaired escape response. PHOX2B is known to be highly express in other types of neurons maintained after ALS progression. Collectively, TARDBP mutations induced loss of axonal resilience, which is an important ALS-related phenotype mediated by PHOX2B downregulation.
A novel deletion in the C-terminal region of HSPB8 in a family with rimmed vacuolar myopathy. International-journal

Aya Inoue-Shibui, Tetsuya Niihori, Michio Kobayashi, Naoki Suzuki, Rumiko Izumi, Hitoshi Warita, Kenju Hara, Matsuyuki Shirota, Ryo Funayama, Keiko Nakayama, Ichizo Nishino, Masashi Aoki, Yoko Aoki

Journal of human genetics　66　(10)　965-972　2021/03/20

DOI： 10.1038/s10038-021-00916-y 　

More details Close

Heat shock protein family B member 8, encoded by HSPB8, is an essential component of the chaperone-assisted selective autophagy complex, which maintains muscle function by degrading damaged proteins in the cells. Mutations in HSPB8 have been reported to cause Charcot-Marie-Tooth type 2L, distal hereditary motor neuropathy IIa, and rimmed vacuolar myopathies (RVM). In this study, we identified a novel heterozygous frameshift variant c.525_529del in HSPB8 in a large Japanese family with RVM, using whole exome sequencing. Three affected individuals had severe respiratory failure, which has not been addressed by previous studies. Muscle atrophy in the paraspinal muscles was also a clinical feature of the individuals affected with RVM in this study. The frameshift mutation was located in the last coding exon, and the mutated protein was predicted to harbor an isoleucine-leucine-valine (ILV) sequence, which corresponds to the IXI/V (isoleucine, X amino acids, and isoleucine or valine) motif. The IXI/V motif is essential for assembly into larger oligomers in other small heat shock proteins and all frameshift mutants of HSPB8 were predicted to share the ILV sequence in the C-terminal extension. The in silico prediction tools showed low protein solubility and increased aggregation propensity for the region around the ILV sequence. The IXI/V motif might be associated with the pathogenesis of HSPB8-related RVM.
Senolysis by glutaminolysis inhibition ameliorates various age-associated disorders. International-journal Peer-reviewed

Johmura Y, Yamanaka T, Omori S, Wang T, Sugiura Y, Matsumoto M, Suzuki N, Kumamoto S, Yamaguchi K, Hatakeyama S, Takami T, Yamaguchi R, Shimizu E, Ikeda K, Okahashi N, Mikawa R, Suematsu M, Arita M, Sugimoto M, Nakayama K, Furukawa Y, Imoto S, Nakanishi M

Science　371　(6526)　265-270　2021/01/15

DOI： 10.1126/science.abb5916 　

More details Close

Removal of senescent cells (senolysis) has been proposed to be beneficial for improving age-associated pathologies, but the molecular pathways for such senolytic activity have not yet emerged. Here, we identified glutaminase 1 (GLS1) as an essential gene for the survival of human senescent cells. The intracellular pH in senescent cells was lowered by lysosomal membrane damage, and this lowered pH induced kidney-type glutaminase (KGA) expression. The resulting enhanced glutaminolysis induced ammonia production, which neutralized the lower pH and improved survival of the senescent cells. Inhibition of KGA-dependent glutaminolysis in aged mice eliminated senescent cells specifically and ameliorated age-associated organ dysfunction. Our results suggest that senescent cells rely on glutaminolysis, and its inhibition offers a promising strategy for inducing senolysis in vivo.
Chromatin Protein PC4 Orchestrates B Cell Differentiation by Collaborating with IKAROS and IRF4. International-journal

Kyoko Ochiai, Mari Yamaoka, Amrutha Swaminathan, Hiroki Shima, Hitoshi Hiura, Mitsuyo Matsumoto, Daisuke Kurotaki, Jun Nakabayashi, Ryo Funayama, Keiko Nakayama, Takahiro Arima, Tomokatsu Ikawa, Tomohiko Tamura, Roger Sciammas, Philippe Bouvet, Tapas K Kundu, Kazuhiko Igarashi

Cell reports　33　(12)　108517-108517　2020/12/22

DOI： 10.1016/j.celrep.2020.108517 　

More details Close

The chromatin protein positive coactivator 4 (PC4) has multiple functions, including chromatin compaction. However, its role in immune cells is largely unknown. We show that PC4 orchestrates chromatin structure and gene expression in mature B cells. B-cell-specific PC4-deficient mice show impaired production of antibody upon antigen stimulation. The PC4 complex purified from B cells contains the transcription factors (TFs) IKAROS and IRF4. IKAROS protein is reduced in PC4-deficient mature B cells, resulting in de-repression of their target genes in part by diminished interactions with gene-silencing components. Upon activation, the amount of IRF4 protein is not increased in PC4-deficient B cells, resulting in reduction of plasma cells. Importantly, IRF4 reciprocally induces PC4 expression via a super-enhancer. PC4 knockdown in human B cell lymphoma and myeloma cells reduces IKAROS protein as an anticancer drug, lenalidomide. Our findings establish PC4 as a chromatin regulator of B cells and a possible therapeutic target adjoining IKAROS in B cell malignancies.
がん研究における女性研究者(WSCR) 大腸がん患者から学んだこと

中山啓子, 舟山亮, 望月保志, 小林実

日本癌学会総会記事　79回　SS2-6　2020/10
Publisher: (一社)日本癌学会
ISSN： 0546-0476
An Amyotrophic Lateral Sclerosis-Associated Mutant of C21ORF2 Is Stabilized by NEK1-Mediated Hyperphosphorylation and the Inability to Bind FBXO3. International-journal

Yasuaki Watanabe, Tadashi Nakagawa, Tetsuya Akiyama, Makiko Nakagawa, Naoki Suzuki, Hitoshi Warita, Masashi Aoki, Keiko Nakayama

iScience　23　(9)　101491-101491　2020/08/21

DOI： 10.1016/j.isci.2020.101491 　

More details Close

C21ORF2 and NEK1 have been identified as amyotrophic lateral sclerosis (ALS)-associated genes. Both genes are also mutated in certain ciliopathies, suggesting that they might contribute to the same signaling pathways. Here we show that FBXO3, the substrate receptor of an SCF ubiquitin ligase complex, binds and ubiquitylates C21ORF2, thereby targeting it for proteasomal degradation. C21ORF2 stabilizes the kinase NEK1, with the result that loss of FBXO3 stabilizes not only C21ORF2 but also NEK1. Conversely, NEK1-mediated phosphorylation stabilizes C21ORF2 by attenuating its interaction with FBXO3. We found that the ALS-associated V58L mutant of C21ORF2 is more susceptible to phosphorylation by NEK1, with the result that it is not ubiquitylated by FBXO3 and therefore accumulates together with NEK1. Expression of C21ORF2(V58L) in motor neurons induced from mouse embryonic stem cells impaired neurite outgrowth. We suggest that inhibition of NEK1 activity is a potential therapeutic approach to ALS associated with C21ORF2 mutation.
Correction: Biallelic GALM pathogenic variants cause a novel type of galactosemia. International-journal

Yoichi Wada, Atsuo Kikuchi, Natsuko Arai-Ichinoi, Osamu Sakamoto, Yusuke Takezawa, Shinya Iwasawa, Tetsuya Niihori, Hiromi Nyuzuki, Yoko Nakajima, Erika Ogawa, Mika Ishige, Hiroki Hirai, Hideo Sasai, Ryoji Fujiki, Matsuyuki Shirota, Ryo Funayama, Masayuki Yamamoto, Tetsuya Ito, Osamu Ohara, Keiko Nakayama, Yoko Aoki, Seizo Koshiba, Toshiyuki Fukao, Shigeo Kure

Genetics in medicine : official journal of the American College of Medical Genetics　22　(7)　1281-1281　2020/07

DOI： 10.1038/s41436-020-0836-z 　

More details Close

An amendment to this paper has been published and can be accessed via a link at the top of the paper.
Variants That Affect Function of Calcium Channel TRPV6 Are Associated With Early-Onset Chronic Pancreatitis. International-journal Peer-reviewed

Atsushi Masamune, Hiroshi Kotani, Franziska Lena Sörgel, Jian-Min Chen, Shin Hamada, Reiko Sakaguchi, Emmanuelle Masson, Eriko Nakano, Yoichi Kakuta, Tetsuya Niihori, Ryo Funayama, Matsuyuki Shirota, Tatsuya Hirano, Tetsuya Kawamoto, Atsuki Hosokoshi, Kiyoshi Kume, Lara Unger, Maren Ewers, Helmut Laumen, Peter Bugert, Masayuki X Mori, Volodymyr Tsvilovskyy, Petra Weißgerber, Ulrich Kriebs, Claudia Fecher-Trost, Marc Freichel, Kalliope N Diakopoulos, Alexandra Berninger, Marina Lesina, Kentaro Ishii, Takao Itoi, Tsukasa Ikeura, Kazuichi Okazaki, Tom Kaune, Jonas Rosendahl, Masao Nagasaki, Yasuhito Uezono, Hana Algül, Keiko Nakayama, Yoichi Matsubara, Yoko Aoki, Claude Férec, Yasuo Mori, Heiko Witt, Tooru Shimosegawa

Gastroenterology　158　(6)　1626-1641　2020/05

DOI： 10.1053/j.gastro.2020.01.005 　

More details Close

BACKGROUND & AIMS: Changes in pancreatic calcium levels affect secretion and might be involved in development of chronic pancreatitis (CP). We investigated the association of CP with the transient receptor potential cation channel subfamily V member 6 gene (TRPV6), which encodes a Ca2+-selective ion channel, in an international cohort of patients and in mice. METHODS: We performed whole-exome DNA sequencing from a patient with idiopathic CP and from his parents, who did not have CP. We validated our findings by sequencing DNA from 300 patients with CP (not associated with alcohol consumption) and 1070 persons from the general population in Japan (control individuals). In replication studies, we sequenced DNA from patients with early-onset CP (20 years or younger) not associated with alcohol consumption from France (n = 470) and Germany (n = 410). We expressed TRPV6 variants in HEK293 cells and measured their activity using Ca2+ imaging assays. CP was induced by repeated injections of cerulein in TRPV6mut/mut mice. RESULTS: We identified the variants c.629C>T (p.A210V) and c.970G>A (p.D324N) in TRPV6 in the index patient. Variants that affected function of the TRPV6 product were found in 13 of 300 patients (4.3%) and 1 of 1070 control individuals (0.1%) from Japan (odds ratio [OR], 48.4; 95% confidence interval [CI], 6.3-371.7; P = 2.4 × 10-8). Twelve of 124 patients (9.7%) with early-onset CP had such variants. In the replication set from Europe, 18 patients with CP (2.0%) carried variants that affected the function of the TRPV6 product compared with 0 control individuals (P = 6.2 × 10-8). Variants that did not affect the function of the TRPV6 product (p.I223T and p.D324N) were overrepresented in Japanese patients vs control individuals (OR, 10.9; 95% CI, 4.5-25.9; P = 7.4 × 10-9 for p.I223T and P = .01 for p.D324N), whereas the p.L299Q was overrepresented in European patients vs control individuals (OR, 3.0; 95% CI, 1.9-4.8; P = 1.2 × 10-5). TRPV6mut/mut mice given cerulein developed more severe pancreatitis than control mice, as shown by increased levels of pancreatic enzymes, histologic alterations, and pancreatic fibrosis. CONCLUSIONS: We found that patients with early-onset CP not associated with alcohol consumption carry variants in TRPV6 that affect the function of its product, perhaps by altering Ca2+ balance in pancreatic cells. TRPV6 regulates Ca2+ homeostasis and pancreatic inflammation.
Longitudinal plasma amino acid profiling with maternal genomic background throughout human pregnancy International-journal Peer-reviewed

Matsuyuki Shirota, Daisuke Saigusa, Riu Yamashita, Yasutake Kato, Mitsuyo Matsumoto, Junya Yamagishi, Noriko Ishida, Kazuki Kumada, Yuji Oe, Hisaaki Kudo, Junji Yokozawa, Yoko Kuroki, Ikuko Motoike, Fumiki Katsuoka, Masao Nagasaki, Seizo Koshiba, Keiko Nakayama, Osamu Tanabe, Jun Yasuda, Shigeo Kure, Kengo Kinoshita, Hirohito Metoki, Shinichi Kuriyama, Nobuo Yaegashi, Masayuki Yamamoto, Junichi Sugawara

Medical Mass Spectrometry　4　(1)　36-49　2020/04/16

DOI： 10.24508/mms.2020.06.001 　
The Autism-Related Protein SETD5 Controls Neural Cell Proliferation through Epigenetic Regulation of rDNA Expression. International-journal Peer-reviewed

Tadashi Nakagawa, Satoko Hattori, Risa Nobuta, Ryuichi Kimura, Makiko Nakagawa, Masaki Matsumoto, Yuko Nagasawa, Ryo Funayama, Tsuyoshi Miyakawa, Toshifumi Inada, Noriko Osumi, Keiichi I Nakayama, Keiko Nakayama

iScience　23　(4)　101030-101030　2020/04/06

DOI： 10.1016/j.isci.2020.101030 　

More details Close

Haploinsufficiency of SETD5 is implicated in syndromic autism spectrum disorder (ASD), but the molecular mechanism underlying the pathological role of this protein has remained unclear. We have now shown that Setd5+/- mice manifest ASD-related behavioral phenotypes and that the expression of ribosomal protein genes and rDNA is disturbed in the brain of these mice. SETD5 recruited the HDAC3 complex to the rDNA promoter, resulting in removal of the histone mark H4K16ac and its reader protein TIP5, a repressor of rDNA expression. Depletion of SETD5 attenuated rDNA expression, translational activity, and neural cell proliferation, whereas ablation of TIP5 in SETD5-deficient cells rescued these effects. Translation of cyclin D1 mRNA was specifically down-regulated in SETD5-insufficient cells. Our results thus suggest that SETD5 positively regulates rDNA expression via an HDAC3-mediated epigenetic mechanism and that such regulation is essential for translation of cyclin D1 mRNA and neural cell proliferation.
BACH1 Promotes Pancreatic Cancer Metastasis by Repressing Epithelial Genes and Enhancing Epithelial-Mesenchymal Transition. International-journal Peer-reviewed

Masaki Sato, Mitsuyo Matsumoto, Yuriko Saiki, Mahabub Alam, Hironari Nishizawa, Masahiro Rokugo, Andrey Brydun, Shinji Yamada, Mika K Kaneko, Ryo Funayama, Mamoru Ito, Yukinari Kato, Keiko Nakayama, Michiaki Unno, Kazuhiko Igarashi

Cancer research　80　(6)　1279-1292　2020/03/15

DOI： 10.1158/0008-5472.CAN-18-4099 　

More details Close

Pancreatic ductal adenocarcinoma (PDAC) is among the cancers with the poorest prognoses due to its highly malignant features. BTB and CNC homology 1 (BACH1) has been implicated in RAS-driven tumor formation. We focused on the role of BACH1 in PDAC, more than 90% of which have KRAS mutation. Knockdown of BACH1 in PDAC cell lines reduced cell migration and invasion, in part, by increasing E-cadherin expression, whereas its overexpression showed opposite effects. BACH1 directly repressed the expression of FOXA1 that is known to activate the expression of CDH1 encoding E-cadherin and to inhibit epithelial-to-mesenchymal transition. BACH1 also directly repressed the expression of genes important for epithelial cell adhesion including CLDN3 and CLDN4. In a mouse orthotopic implantation model, BACH1 was required for the high metastatic ability of AsPC-1 cells. IHC analysis of clinical specimens with a newly developed anti-BACH1 mAb revealed that high expression of BACH1 is a poor prognostic factor. These results suggest that the gene regulatory network of BACH1 and downstream genes including CDH1 contribute to the malignant features of PDAC by regulating epithelial-to-mesenchymal transition. SIGNIFICANCE: Greater understanding of the gene regulatory network involved in epithelial-to-mesenchymal transition of pancreatic cancer cells will provide novel therapeutic targets and diagnostic markers.
Metabolic and pathologic profiles of human LSS deficiency recapitulated in mice. International-journal Peer-reviewed

Yoichi Wada, Atsuo Kikuchi, Akimune Kaga, Naoki Shimizu, Junya Ito, Ryo Onuma, Fumiyoshi Fujishima, Eriko Totsune, Ryo Sato, Tetsuya Niihori, Matsuyuki Shirota, Ryo Funayama, Kota Sato, Toru Nakazawa, Keiko Nakayama, Yoko Aoki, Setsuya Aiba, Kiyotaka Nakagawa, Shigeo Kure

PLoS genetics　16　(2)　e1008628　2020/02

DOI： 10.1371/journal.pgen.1008628 　

More details Close

Skin lesions, cataracts, and congenital anomalies have been frequently associated with inherited deficiencies in enzymes that synthesize cholesterol. Lanosterol synthase (LSS) converts (S)-2,3-epoxysqualene to lanosterol in the cholesterol biosynthesis pathway. Biallelic mutations in LSS have been reported in families with congenital cataracts and, very recently, have been reported in cases of hypotrichosis. However, it remains to be clarified whether these phenotypes are caused by LSS enzymatic deficiencies in each tissue, and disruption of LSS enzymatic activity in vivo has not yet been validated. We identified two patients with novel biallelic LSS mutations who exhibited congenital hypotrichosis and midline anomalies but did not have cataracts. We showed that the blockade of the LSS enzyme reaction occurred in the patients by measuring the (S)-2,3-epoxysqualene/lanosterol ratio in the forehead sebum, which would be a good biomarker for the diagnosis of LSS deficiency. Epidermis-specific Lss knockout mice showed neonatal lethality due to dehydration, indicating that LSS could be involved in skin barrier integrity. Tamoxifen-induced knockout of Lss in the epidermis caused hypotrichosis in adult mice. Lens-specific Lss knockout mice had cataracts. These results confirmed that LSS deficiency causes hypotrichosis and cataracts due to loss-of-function mutations in LSS in each tissue. These mouse models will lead to the elucidation of the pathophysiological mechanisms associated with disrupted LSS and to the development of therapeutic treatments for LSS deficiency.
Cohort Profile: Tohoku Medical Megabank Project Birth and Three-Generation Cohort Study (TMM BirThree Cohort Study): rationale, progress and perspective. International-journal Peer-reviewed

Shinichi Kuriyama, Hirohito Metoki, Masahiro Kikuya, Taku Obara, Mami Ishikuro, Chizuru Yamanaka, Masato Nagai, Hiroko Matsubara, Tomoko Kobayashi, Junichi Sugawara, Gen Tamiya, Atsushi Hozawa, Naoki Nakaya, Naho Tsuchiya, Tomohiro Nakamura, Akira Narita, Mana Kogure, Takumi Hirata, Ichiro Tsuji, Fuji Nagami, Nobuo Fuse, Tomohiko Arai, Yoshio Kawaguchi, Shinichi Higuchi, Masaki Sakaida, Yoichi Suzuki, Noriko Osumi, Keiko Nakayama, Kiyoshi Ito, Shinichi Egawa, Koichi Chida, Eiichi Kodama, Hideyasu Kiyomoto, Tadashi Ishii, Akito Tsuboi, Hiroaki Tomita, Yasuyuki Taki, Hiroshi Kawame, Kichiya Suzuki, Naoto Ishii, Soichi Ogishima, Satoshi Mizuno, Takako Takai-Igarashi, Naoko Minegishi, Jun Yasuda, Kazuhiko Igarashi, Ritsuko Shimizu, Masao Nagasaki, Osamu Tanabe, Seizo Koshiba, Hiroaki Hashizume, Hozumi Motohashi, Teiji Tominaga, Sadayoshi Ito, Kozo Tanno, Kiyomi Sakata, Atsushi Shimizu, Jiro Hitomi, Makoto Sasaki, Kengo Kinoshita, Hiroshi Tanaka, Tadao Kobayashi, Shigeo Kure, Nobuo Yaegashi, Masayuki Yamamoto

International journal of epidemiology　49　(1)　18-19　2020/02/01

DOI： 10.1093/ije/dyz169 　

ISSN： 0300-5771
Study profile of The Tohoku Medical Megabank Community-Based Cohort Study. Peer-reviewed

Atsushi Hozawa, Kozo Tanno, Naoki Nakaya, Tomohiro Nakamura, Naho Tsuchiya, Takumi Hirata, Akira Narita, Mana Kogure, Kotaro Nochioka, Ryohei Sasaki, Nobuyuki Takanashi, Kotaro Otsuka, Kiyomi Sakata, Shinichi Kuriyama, Masahiro Kikuya, Osamu Tanabe, Junichi Sugawara, Kichiya Suzuki, Yoichi Suzuki, Eiichi N Kodama, Nobuo Fuse, Hideyasu Kiyomoto, Hiroaki Tomita, Akira Uruno, Yohei Hamanaka, Hirohito Metoki, Mami Ishikuro, Taku Obara, Tomoko Kobayashi, Kazuyuki Kitatani, Takako Takai-Igarashi, Soichi Ogishima, Mamoru Satoh, Hideki Ohmomo, Akito Tsuboi, Shinichi Egawa, Tadashi Ishii, Kiyoshi Ito, Sadayoshi Ito, Yasuyuki Taki, Naoko Minegishi, Naoto Ishii, Masao Nagasaki, Kazuhiko Igarashi, Seizo Koshiba, Ritsuko Shimizu, Gen Tamiya, Keiko Nakayama, Hozumi Motohashi, Jun Yasuda, Atsushi Shimizu, Tsuyoshi Hachiya, Yuh Shiwa, Teiji Tominaga, Hiroshi Tanaka, Kotaro Oyama, Ryoichi Tanaka, Hiroshi Kawame, Akimune Fukushima, Yasushi Ishigaki, Tomoharu Tokutomi, Noriko Osumi, Tadao Kobayashi, Fuji Nagami, Hiroaki Hashizume, Tomohiro Arai, Yoshio Kawaguchi, Shinichi Higuchi, Masaki Sakaida, Ryujin Endo, Satoshi Nishizuka, Ichiro Tsuji, Jiro Hitomi, Motoyuki Nakamura, Kuniaki Ogasawara, Nobuo Yaegashi, Kengo Kinoshita, Shigeo Kure, Akio Sakai, Seiichiro Kobayashi, Kenji Sobue, Makoto Sasaki, Masayuki Yamamoto

Journal of epidemiology　31　(1)　65-76　2020/01/11

DOI： 10.2188/jea.JE20190271 　

More details Close

BackgroundWe established a community-based cohort study to assess the long-term impact of the Great East Japan Earthquake on disaster victims and gene-environmental interactions on the incidence of major diseases such as cancer and cardiovascular diseases.MethodsWe asked participants to join our cohort in the health check-up settings and assessment center based settings. Inclusion criteria was aged 20 years or over and living in Miyagi or Iwate Prefecture. We obtained information on lifestyle, effect of disaster, blood, and urine information (Type 1 survey), and some detailed measurements (Type 2 survey), for example, carotid echography, calcaneal ultrasound bone mineral density, and so on. All participants agreed to measure genome information and to distribute their information widely.ResultsAs a result, 87,865 gave their informed consent to join our study. Participation rate at health check-up site was about 70%. The participants with Type 1 survey were more likely to have psychological distress than those of Type 2 survey, and women were more likely to have psychological distress than men. Additionally, coastal residents were more likely to have higher degrees of psychological distress than inland residents regardless of sex.ConclusionThis cohort comprised large sample size and it contains information on disaster, genome information, and metabolome information. This cohort also had several detailed measurements. Using this cohort enabled us to clarify the long-term effect of disaster and also to establish personalized prevention based on genome, metabolome, and other omics information.
Knockout Mouse Models Provide Insight into the Biological Functions of CRL1 Components. International-journal Peer-reviewed

Tadashi Nakagawa, Keiko Nakayama, Keiichi I Nakayama

Advances in experimental medicine and biology　1217　147-171　2020

DOI： 10.1007/978-981-15-1025-0_10 　

More details Close

The CRL1 complex, also known as the SCF complex, is a ubiquitin ligase that in mammals consists of an adaptor protein (SKP1), a scaffold protein (CUL1), a RING finger protein (RBX1, also known as ROC1), and one of about 70 F-box proteins. Given that the F-box proteins determine the substrate specificity of the CRL1 complex, the variety of these proteins allows the generation of a large number of ubiquitin ligases that promote the degradation or regulate the function of many substrate proteins and thereby control numerous key cellular processes. The physiological and pathological functions of these many CRL1 ubiquitin ligases have been studied by the generation and characterization of knockout mouse models that lack specific CRL1 components. In this chapter, we provide a comprehensive overview of these mouse models and discuss the role of each CRL1 component in mouse physiology and pathology.
Pathogenic mutations in the ALS gene CCNF cause cytoplasmic mislocalization of Cyclin F and elevated VCP ATPase activity. International-journal Peer-reviewed

Yujiao Yu, Tadashi Nakagawa, Akane Morohoshi, Makiko Nakagawa, Noriko Ishida, Naoki Suzuki, Masashi Aoki, Keiko Nakayama

Human molecular genetics　28　(20)　3486-3497　2019/10/15

DOI： 10.1093/hmg/ddz119 　

ISSN： 0964-6906

More details Close

Amyotrophic lateral sclerosis (ALS) is an adult-onset motor neuron disease characterized by a progressive decline in motor function. Genetic analyses have identified several genes mutated in ALS patients, and one of them is Cyclin F gene (CCNF), the product of which (Cyclin F) serves as the substrate-binding module of a SKP1-CUL1-F-box protein (SCF) ubiquitin ligase complex. However, the role of Cyclin F in ALS pathogenesis has remained unclear. Here, we show that Cyclin F binds to valosin-containing protein (VCP), which is also reported to be mutated in ALS, and that the two proteins colocalize in the nucleus. VCP was found to bind to the NH2-terminal region of Cyclin F and was not ubiquitylated by SCFCyclin F in transfected cells. Instead, the ATPase activity of VCP was enhanced by Cyclin F in vitro. Furthermore, whereas ALS-associated mutations of CCNF did not affect the stability of Cyclin F or disrupt formation of the SCFCyclin F complex, amino acid substitutions in the VCP binding region increased the binding ability of Cyclin F to VCP and activity of VCP as well as mislocalization of the protein in the cytoplasm. We also provided evidence that the ATPase activity of VCP promotes cytoplasmic aggregation of transactivation responsive region (TAR) DNA-binding protein 43, which is commonly observed in degenerating neurons in ALS patients. Given that mutations of VCP identified in ALS patients also increase its ATPase activity, our results suggest that Cyclin F mutations may contribute to ALS pathogenesis by increasing the ATPase activity of VCP in the cytoplasm, which in turn increases TDP-43 aggregates.
Cancer-associated fibroblasts secrete Wnt2 to promote cancer progression in colorectal cancer. International-journal Peer-reviewed

Takashi Aizawa, Hideaki Karasawa, Ryo Funayama, Matsuyuki Shirota, Takashi Suzuki, Shimpei Maeda, Hideyuki Suzuki, Akihiro Yamamura, Takeshi Naitoh, Keiko Nakayama, Michiaki Unno

Cancer medicine　8　(14)　6370-6382　2019/10

DOI： 10.1002/cam4.2523 　

More details Close

Recent studies have shown that the tumor microenvironment plays a significant role in the progression of solid tumors. As an abundant component of the tumor microenvironment, cancer-associated fibroblasts (CAFs) have been shown to promote tumorigenesis and cancer aggressiveness, but their molecular characteristics remain poorly understood. In the present study, paired CAFs and normal fibroblasts (NFs) were isolated from five colorectal cancer (CRC) tissues from patients who underwent surgical resection. The gene expression profiles of CAFs and NFs identified by RNA sequencing were compared to understand the complex role of CAFs in cancer progression. Gene Set Enrichment Analysis revealed that the gene sets related to the Wnt signaling pathway were highly enriched in CAFs, as well as TGFβ signaling, which is considered to be a regulator of CAFs. Among the components of this pathway, Wnt2 was specifically expressed. The observations led us to speculate that Wnt2 is extremely involved in regulating CRC progression by CAFs. Thus, we performed immunohistochemical analysis on Wnt2 in 171 patients who underwent surgery for colorectal adenocarcinoma. Positive staining for Wnt2 was mainly observed in cancer stroma, although the immunoreactivity was weak in cancer cells. Wnt2 expression in CAFs was significantly correlated with depth of tumor (P < .001), lymph node metastasis (P = .044), TNM stage (P = .010), venous invasion (P < .001), and recurrence (P = .013). Subsequent in vitro analyses were conducted using conditioned medium (CM) from immortalized CAFs transfected with siRNA targeting Wnt2. As a result, cancer cell invasion and migration were significantly decreased in the CM from immortalized CAFs transfected with siRNA targeting Wnt2. Our findings indicated that Wnt2 protein released from CAFs enhances CRC cell invasion and migration. In conclusion, Wnt2 secreted by CAFs plays a key role in cancer progression and is a potential therapeutic target for CRC.
Detection of NRAS mutation in cell-free DNA biological fluids from patients with kaposiform lymphangiomatosis. International-journal Peer-reviewed

Michio Ozeki, Yoko Aoki, Akifumi Nozawa, Shiho Yasue, Saori Endo, Yumiko Hori, Kentaro Matsuoka, Tetsuya Niihori, Ryo Funayama, Matsuyuki Shirota, Keiko Nakayama, Toshiyuki Fukao

Orphanet journal of rare diseases　14　(1)　215-215　2019/09/11

DOI： 10.1186/s13023-019-1191-5 　

More details Close

BACKGROUND: Kaposiform lymphangiomatosis (KLA) has recently been distinguished as a novel subtype of generalized lymphatic anomaly (GLA) with foci of spindle endothelial cells. All cases of KLA involve multiple organs and have an unfavorable prognosis. However, the molecular pathogenesis is unknown, and there are no useful biomarkers. In the present study, we performed genetic analysis to elucidate the cause of this disease and detect biomarkers for it. METHODS: We performed whole-exome sequencing of DNA samples from leukocytes and a biopsy specimen and analyzed cell-free DNA (cfDNA) from plasma and pleural effusion of patients to identify the NRAS c.182A > G (p.Q61R) mutation using the droplet digital polymerase chain reaction (ddPCR). RESULTS: All KLA patients (patients 1-5) had invasive and aggressive features (hemorrhagic pleural effusions, coagulation disorder, and thrombocytopenia) and characteristic findings of KLA in their pathological examinations. In whole exome sequencing for patient 1, c.182A > G missense variant (p.Q61R) in NRAS was identified in fresh frozen samples of a mass on the left chest wall at a frequency of 5% of total alleles but not in his blood leukocytes. Furthermore, the same mutation was detected in cfDNA isolated from plasma and pleural effusion by using ddPCR. ddPCR analysis of plasma/pleural effusion samples from an additional four KLA patients showed that the same mutation was detected in isolated cfDNA in three of the four, as well as in a tissue sample from one of the three plasma/effusion-positive patients that had been obtained to confirm the mutation. CONCLUSION: These results provide the first evidence that NRAS oncogenic variant was identified in DNA samples from KLA patients from not only two affected lesions but also plasma and pleural effusion.
ペルオキシソーム形成異常による小脳形態形成障害の病態発症機構

阿部雄一, 本庄雅則, 伊藤竜太, 藤谷昌司, 中山啓子, 大城遼平, 中山敬一, 山下俊英, 藤木幸夫

日本生化学会大会プログラム・講演要旨集　92回　[1T04a-06]　2019/09
Publisher: (公社)日本生化学会
Aberrant axon branching via Fos-B dysregulation in FUS-ALS motor neurons. International-journal Peer-reviewed

Tetsuya Akiyama, Naoki Suzuki, Mitsuru Ishikawa, Koki Fujimori, Takefumi Sone, Jiro Kawada, Ryo Funayama, Fumiyoshi Fujishima, Shio Mitsuzawa, Kensuke Ikeda, Hiroya Ono, Tomomi Shijo, Shion Osana, Matsuyuki Shirota, Tadashi Nakagawa, Yasuo Kitajima, Ayumi Nishiyama, Rumiko Izumi, Satoru Morimoto, Yohei Okada, Takayuki Kamei, Mayumi Nishida, Masahiro Nogami, Shohei Kaneda, Yoshiho Ikeuchi, Hiroaki Mitsuhashi, Keiko Nakayama, Teruo Fujii, Hitoshi Warita, Hideyuki Okano, Masashi Aoki

EBioMedicine　45　362-378　2019/07

DOI： 10.1016/j.ebiom.2019.06.013 　

More details Close

BACKGROUND: The characteristic structure of motor neurons (MNs), particularly of the long axons, becomes damaged in the early stages of amyotrophic lateral sclerosis (ALS). However, the molecular pathophysiology of axonal degeneration remains to be fully elucidated. METHOD: Two sets of isogenic human-induced pluripotent stem cell (hiPSCs)-derived MNs possessing the single amino acid difference (p.H517D) in the fused in sarcoma (FUS) were constructed. By combining MN reporter lentivirus, MN specific phenotype was analyzed. Moreover, RNA profiling of isolated axons were conducted by applying the microfluidic devices that enable axon bundles to be produced for omics analysis. The relationship between the target gene, which was identified as a pathological candidate in ALS with RNA-sequencing, and the MN phenotype was confirmed by intervention with si-RNA or overexpression to hiPSCs-derived MNs and even in vivo. The commonality was further confirmed with other ALS-causative mutant hiPSCs-derived MNs and human pathology. FINDINGS: We identified aberrant increasing of axon branchings in FUS-mutant hiPSCs-derived MN axons compared with isogenic controls as a novel phenotype. We identified increased level of Fos-B mRNA, the binding target of FUS, in FUS-mutant MNs. While Fos-B reduction using si-RNA or an inhibitor ameliorated the observed aberrant axon branching, Fos-B overexpression resulted in aberrant axon branching even in vivo. The commonality of those phenotypes was further confirmed with other ALS causative mutation than FUS. INTERPRETATION: Analyzing the axonal fraction of hiPSC-derived MNs using microfluidic devices revealed that Fos-B is a key regulator of FUS-mutant axon branching. FUND: Japan Agency for Medical Research and development; Japanese Ministry of Education, Culture, Sports, Science and Technology Clinical Research, Innovation and Education Center, Tohoku University Hospital; Japan Intractable Diseases (Nanbyo) Research Foundation; the Kanae Foundation for the Promotion of Medical Science; and "Inochi-no-Iro" ALS research grant.
Germline-Activating RRAS2 Mutations Cause Noonan Syndrome. International-journal Peer-reviewed

Tetsuya Niihori, Koki Nagai, Atsushi Fujita, Hirofumi Ohashi, Nobuhiko Okamoto, Satoshi Okada, Atsuko Harada, Hirotaka Kihara, Thomas Arbogast, Ryo Funayama, Matsuyuki Shirota, Keiko Nakayama, Taiki Abe, Shin-Ichi Inoue, I-Chun Tsai, Naomichi Matsumoto, Erica E Davis, Nicholas Katsanis, Yoko Aoki

American journal of human genetics　104　(6)　1233-1240　2019/06/06

DOI： 10.1016/j.ajhg.2019.04.014 　

ISSN： 0002-9297

More details Close

Noonan syndrome (NS) is characterized by distinctive craniofacial appearance, short stature, and congenital heart disease. Approximately 80% of individuals with NS harbor mutations in genes whose products are involved in the RAS/mitogen-activating protein kinase (MAPK) pathway. However, the underlying genetic causes in nearly 20% of individuals with NS phenotype remain unexplained. Here, we report four de novo RRAS2 variants in three individuals with NS. RRAS2 is a member of the RAS subfamily and is ubiquitously expressed. Three variants, c.70_78dup (p.Gly24_Gly26dup), c.216A>T (p.Gln72His), and c.215A>T (p.Gln72Leu), have been found in cancers; our functional analyses showed that these three changes induced elevated association of RAF1 and that they activated ERK1/2 and ELK1. Notably, prominent activation of ERK1/2 and ELK1 by p.Gln72Leu associates with the severe phenotype of the individual harboring this change. To examine variant pathogenicity in vivo, we generated zebrafish models. Larvae overexpressing c.70_78dup (p.Gly24_Gly26dup) or c.216A>T (p.Gln72His) variants, but not wild-type RRAS2 RNAs, showed craniofacial defects and macrocephaly. The same dose injection of mRNA encoding c.215A>T (p.Gln72Leu) caused severe developmental impairments and low dose overexpression of this variant induced craniofacial defects. In contrast, the RRAS2 c.224T>G (p.Phe75Cys) change, located on the same allele with p.Gln72His in an individual with NS, resulted in no aberrant in vitro or in vivo phenotypes by itself. Together, our findings suggest that activating RRAS2 mutations can cause NS and expand the involvement of RRAS2 proto-oncogene to rare germline disorders.
Biallelic GALM pathogenic variants cause a novel type of galactosemia. International-journal Peer-reviewed

Yoichi Wada, Atsuo Kikuchi, Natsuko Arai-Ichinoi, Osamu Sakamoto, Yusuke Takezawa, Shinya Iwasawa, Tetsuya Niihori, Hiromi Nyuzuki, Yoko Nakajima, Erika Ogawa, Mika Ishige, Hiroki Hirai, Hideo Sasai, Ryoji Fujiki, Matsuyuki Shirota, Ryo Funayama, Masayuki Yamamoto, Tetsuya Ito, Osamu Ohara, Keiko Nakayama, Yoko Aoki, Seizo Koshiba, Toshiyuki Fukao, Shigeo Kure

Genetics in medicine : official journal of the American College of Medical Genetics　21　(6)　1286-1294　2019/06

DOI： 10.1038/s41436-018-0340-x 　

ISSN： 1098-3600

More details Close

PURPOSE: Galactosemia is caused by metabolic disturbances at various stages of galactose metabolism, including deficiencies in enzymes involved in the Leloir pathway (GALT, GALK1, and GALE). Nevertheless, the etiology of galactosemia has not been identified in a subset of patients. This study aimed to explore the causes of unexplained galactosemia. METHODS: Trio-based exome sequencing and/or Sanger sequencing was performed in eight patients with unexplained congenital galactosemia. In vitro enzymatic assays and immunoblot assays were performed to confirm the pathogenicity of the variants. RESULTS: The highest blood galactose levels observed in each patient were 17.3-41.9 mg/dl. Bilateral cataracts were observed in two patients. In all eight patients, we identified biallelic variants (p.Arg82*, p.Ile99Leufs*46, p.Gly142Arg, p.Arg267Gly, and p.Trp311*) in the GALM encoding galactose mutarotase, which catalyzes epimerization between β- and α-D-galactose in the first step of the Leloir pathway. GALM enzyme activities were undetectable in lymphoblastoid cell lines established from two patients. Immunoblot analysis showed the absence of the GALM protein in the patients' peripheral blood mononuclear cells. In vitro GALM expression and protein stability assays revealed altered stabilities of the variant GALM proteins. CONCLUSION: Biallelic GALM pathogenic variants cause galactosemia, suggesting the existence of type IV galactosemia.
Recurrent de novo MAPK8IP3 variants cause neurological phenotypes. International-journal Peer-reviewed

Shinya Iwasawa, Kumiko Yanagi, Atsuo Kikuchi, Yasuko Kobayashi, Kazuhiro Haginoya, Hiroshi Matsumoto, Kenji Kurosawa, Masayuki Ochiai, Yasunari Sakai, Atsushi Fujita, Noriko Miyake, Tetsuya Niihori, Matsuyuki Shirota, Ryo Funayama, Shigeaki Nonoyama, Shouichi Ohga, Hiroshi Kawame, Keiko Nakayama, Yoko Aoki, Naomichi Matsumoto, Tadashi Kaname, Yoichi Matsubara, Wataru Shoji, Shigeo Kure

Annals of neurology　85　(6)　927-933　2019/06

DOI： 10.1002/ana.25481 　

ISSN： 0364-5134

More details Close

c-Jun-amino-terminal kinase-interacting protein 3 (JIP3), encoded by MAPK8IP3, is an adaptor protein of the kinesin-1 complex and essential for axonal transport in neurons. However, an association between MAPK8IP3 variants and human disease has not been established. We identified 5 individuals from four families with recurrent de novo variants c.1732C>T (p.Arg578Cys) and c.3436C>T (p.Arg1146Cys) in MAPK8IP3. The core phenotype includes spastic diplegia, intellectual disability, cerebral atrophy, and corpus callosum hypoplasia. Zebrafish embryos overexpressing human mutant JIP3 showed axon varicosities of the posterior lateral line nerve, suggesting an adverse effect on the developing axons. Our results suggest that MAPK8IP3 variants cause a neurodevelopmental disease. ANN NEUROL 2019;85:927-933.
遺伝学的分析から特定された17患者中9名における病原性変異の候補(Genomic analysis identified candidate pathogenic variants in 9 of 17 patients)

Takezawa Yusuke, Kikuchi Atsuo, Haginoya Kazuhiro, Niihari Tetsuya, Numata-Uematsu Yurika, Inui Takehiko, Yamamura-Suzuki Saeko, Miyabayashi Takuya, Anzai Mai, Suzuki-Muromoto Sato, Okubo Yukimune, Endo Wakaba, Togashi Noriko, Kobayashi Yasuko, Onuma Akira, Funayama Ryo, Shirota Matsuyuki, Nakayama Keiko, Aoki Yoko, Kure Shigeo

日本小児科学会雑誌　123　(2)　292-292　2019/02
Publisher: (公社)日本小児科学会
ISSN： 0001-6543
Peroxisome biogenesis deficiency attenuates the BDNF-TrkB pathway-mediated development of the cerebellum. International-journal Peer-reviewed

Yuichi Abe, Masanori Honsho, Ryota Itoh, Ryoko Kawaguchi, Masashi Fujitani, Kazushirou Fujiwara, Masaaki Hirokane, Takashi Matsuzaki, Keiko Nakayama, Ryohei Ohgi, Toshihiro Marutani, Keiichi I Nakayama, Toshihide Yamashita, Yukio Fujiki

Life science alliance　1　(6)　e201800062　2018/12

DOI： 10.26508/lsa.201800062 　

More details Close

Peroxisome biogenesis disorders (PBDs) manifest as neurological deficits in the central nervous system, including neuronal migration defects and abnormal cerebellum development. However, the mechanisms underlying pathogenesis remain enigmatic. Here, to investigate how peroxisome deficiency causes neurological defects of PBDs, we established a new PBD model mouse defective in peroxisome assembly factor Pex14p, termed Pex14 ΔC/ΔC mouse. Pex14 ΔC/ΔC mouse manifests a severe symptom such as disorganization of cortical laminar structure and dies shortly after birth, although peroxisomal biogenesis and metabolism are partially defective. The Pex14 ΔC/ΔC mouse also shows malformation of the cerebellum including the impaired dendritic development of Purkinje cells. Moreover, extracellular signal-regulated kinase and AKT signaling are attenuated in this mutant mouse by an elevated level of brain-derived neurotrophic factor (BDNF) together with the enhanced expression of TrkB-T1, a dominant-negative isoform of the BDNF receptor. Our results suggest that dysregulation of the BDNF-TrkB pathway, an essential signaling for cerebellar morphogenesis, gives rise to the pathogenesis of the cerebellum in PBDs.
Novel IARS2 mutations in Japanese siblings with CAGSSS, Leigh, and West syndrome. International-journal Peer-reviewed

Yusuke Takezawa, Hiromi Fujie, Atsuo Kikuchi, Tetsuya Niihori, Ryo Funayama, Matsuyuki Shirota, Keiko Nakayama, Yoko Aoki, Masayuki Sasaki, Shigeo Kure

Brain & development　40　(10)　934-938　2018/11

DOI： 10.1016/j.braindev.2018.06.010 　

ISSN： 0387-7604

More details Close

BACKGROUND: IARS2 encodes isoleucine-tRNA synthetase, which is aclass-1 amino acyl-tRNA synthetase. IARS2 mutations are reported to cause Leigh syndrome or cataracts, growth hormone deficiency, sensory neuropathy, sensorineural hearing loss, and skeletal dysphasia syndrome (CAGSSS). To our knowledge, IARS2 mutations and diseases related to it have only been reported in three families. Here we report a case of two Japanese siblings with Leigh syndrome, some features of CAGSSS, and West syndrome that are found to have compound heterozygous novel IARS2 mutations. CASE REPORT: A 7-month-old Japanese girl presented with infantile spasms. Brain magnetic resonance imaging (MRI) revealed diffuse brain atrophy and hyperintensity in the bilateral basal ganglia. Three years later, her younger sister also presented with infantile spasms. MRI revealed diffuse brain atrophy and hyperintensity of the bilateral ganglia, suggesting Leigh syndrome. The siblings were identified with compound heterozygous missense mutations in IARS2, p.[(Phe227Ser)];[(Arg817His)]. CONCLUSION: This is the first case study reporting Leigh syndrome concomitant with some features of CAGSSS in siblings with novel IARS2 mutations, thereby broadening the phenotypic spectrum of IARS2-related disorders. Further studies are warranted to elucidate the nature of these disorders.
The ubiquitin ligase subunit β-TrCP in Sertoli cells is essential for spermatogenesis in mice. Peer-reviewed

Morohoshi A, Nakagawa T, Nakano S, Nagasawa Y, Nakayama K

Developmental biology　445　(2)　178-188　2018/11

DOI： 10.1016/j.ydbio.2018.10.023 　

ISSN： 0012-1606
Bone marrow PDGFRα+Sca-1+-enriched mesenchymal stem cells support survival of and antibody production by plasma cells in vitro through IL-6 Peer-reviewed

Atsuko Kayaba, Ari Itoh-Nakadai, Kunimichi Niibe, Matsuyuki Shirota, Ryo Funayama, Akiko Sugahara-Tobinai, Yi Li Wong, Masanori Inui, Keiko Nakayama, Toshiyuki Takai

International Immunology　30　(6)　241-253　2018/05/24
Publisher: Oxford University Press
DOI： 10.1093/intimm/dxy018 　

ISSN： 1460-2377 0953-8178
Mutations in six nephrosis genes delineate a pathogenic pathway amenable to treatment. International-journal Peer-reviewed

Shazia Ashraf, Hiroki Kudo, Jia Rao, Atsuo Kikuchi, Eugen Widmeier, Jennifer A Lawson, Weizhen Tan, Tobias Hermle, Jillian K Warejko, Shirlee Shril, Merlin Airik, Tilman Jobst-Schwan, Svjetlana Lovric, Daniela A Braun, Heon Yung Gee, David Schapiro, Amar J Majmundar, Carolin E Sadowski, Werner L Pabst, Ankana Daga, Amelie T van der Ven, Johanna M Schmidt, Boon Chuan Low, Anjali Bansal Gupta, Brajendra K Tripathi, Jenny Wong, Kirk Campbell, Kay Metcalfe, Denny Schanze, Tetsuya Niihori, Hiroshi Kaito, Kandai Nozu, Hiroyasu Tsukaguchi, Ryojiro Tanaka, Kiyoshi Hamahira, Yasuko Kobayashi, Takumi Takizawa, Ryo Funayama, Keiko Nakayama, Yoko Aoki, Naonori Kumagai, Kazumoto Iijima, Henry Fehrenbach, Jameela A Kari, Sherif El Desoky, Sawsan Jalalah, Radovan Bogdanovic, Nataša Stajić, Hildegard Zappel, Assel Rakhmetova, Sharon-Rose Wassmer, Therese Jungraithmayr, Juergen Strehlau, Aravind Selvin Kumar, Arvind Bagga, Neveen A Soliman, Shrikant M Mane, Lewis Kaufman, Douglas R Lowy, Mohamad A Jairajpuri, Richard P Lifton, York Pei, Martin Zenker, Shigeo Kure, Friedhelm Hildebrandt

Nature communications　9　(1)　1960-1960　2018/05/17

DOI： 10.1038/s41467-018-04193-w 　

More details Close

No efficient treatment exists for nephrotic syndrome (NS), a frequent cause of chronic kidney disease. Here we show mutations in six different genes (MAGI2, TNS2, DLC1, CDK20, ITSN1, ITSN2) as causing NS in 17 families with partially treatment-sensitive NS (pTSNS). These proteins interact and we delineate their roles in Rho-like small GTPase (RLSG) activity, and demonstrate deficiency for mutants of pTSNS patients. We find that CDK20 regulates DLC1. Knockdown of MAGI2, DLC1, or CDK20 in cultured podocytes reduces migration rate. Treatment with dexamethasone abolishes RhoA activation by knockdown of DLC1 or CDK20 indicating that steroid treatment in patients with pTSNS and mutations in these genes is mediated by this RLSG module. Furthermore, we discover ITSN1 and ITSN2 as podocytic guanine nucleotide exchange factors for Cdc42. We generate Itsn2-L knockout mice that recapitulate the mild NS phenotype. We, thus, define a functional network of RhoA regulation, thereby revealing potential therapeutic targets.
Rett-like features and cortical visual impairment in a Japanese patient with HECW2 mutation. International-journal Peer-reviewed

Haruhiko Nakamura, Mitsugu Uematsu, Yurika Numata-Uematsu, Yu Abe, Wakaba Endo, Atsuo Kikuchi, Yusuke Takezawa, Ryo Funayama, Matsuyuki Shirota, Keiko Nakayama, Tetsuya Niihori, Yoko Aoki, Kazuhiro Haginoya, Shigeo Kure

Brain & development　40　(5)　410-414　2018/05

DOI： 10.1016/j.braindev.2017.12.015 　

ISSN： 0387-7604

More details Close

Numerous genetic syndromes that include intellectual disability (ID) have been reported. Recently, HECW2 mutations were detected in patients with ID and growth development disorders. Four de novo missense mutations have been reported. Here, we report a Japanese girl with Rett-like symptoms of severe ID, hypotonia, refractory epilepsy, and stereotypical hand movement (hand tapping, flapping, and wringing) after the age of 1 year. Characteristically, she had cortical visual impairment. She had difficulty swallowing since the age of 4 years, and diminished activity was noticeable since the age of 12 years, suggesting neurodevelopmental regression. She has no acquired microcephaly, and brain magnetic resonance imaging showed non-specific mild cerebral and cerebellar atrophy without progression over time. Genetic analyses of MECP2, CDKL5, and FOXG1 were negative. Whole-exome sequencing analysis revealed a known de novo mutation (c.3988C > T) in HECW2. The characteristics of her clinical symptoms are severe cortical visual impairment and Rett-like phenotype such as involuntary movements and regression. This is the first report that patients with HECW2 mutation could show Rett-like feature.
Genomic analysis identifies masqueraders of full-term cerebral palsy. International-journal Peer-reviewed

Yusuke Takezawa, Atsuo Kikuchi, Kazuhiro Haginoya, Tetsuya Niihori, Yurika Numata-Uematsu, Takehiko Inui, Saeko Yamamura-Suzuki, Takuya Miyabayashi, Mai Anzai, Sato Suzuki-Muromoto, Yukimune Okubo, Wakaba Endo, Noriko Togashi, Yasuko Kobayashi, Akira Onuma, Ryo Funayama, Matsuyuki Shirota, Keiko Nakayama, Yoko Aoki, Shigeo Kure

Annals of clinical and translational neurology　5　(5)　538-551　2018/05

DOI： 10.1002/acn3.551 　

More details Close

Objective: Cerebral palsy is a common, heterogeneous neurodevelopmental disorder that causes movement and postural disabilities. Recent studies have suggested genetic diseases can be misdiagnosed as cerebral palsy. We hypothesized that two simple criteria, that is, full-term births and nonspecific brain MRI findings, are keys to extracting masqueraders among cerebral palsy cases due to the following: (1) preterm infants are susceptible to multiple environmental factors and therefore demonstrate an increased risk of cerebral palsy and (2) brain MRI assessment is essential for excluding environmental causes and other particular disorders. Methods: A total of 107 patients-all full-term births-without specific findings on brain MRI were identified among 897 patients diagnosed with cerebral palsy who were followed at our center. DNA samples were available for 17 of the 107 cases for trio whole-exome sequencing and array comparative genomic hybridization. We prioritized variants in genes known to be relevant in neurodevelopmental diseases and evaluated their pathogenicity according to the American College of Medical Genetics guidelines. Results: Pathogenic/likely pathogenic candidate variants were identified in 9 of 17 cases (52.9%) within eight genes: CTNNB1,CYP2U1,SPAST,GNAO1,CACNA1A,AMPD2,STXBP1, and SCN2A. Five identified variants had previously been reported. No pathogenic copy number variations were identified. The AMPD2 missense variant and the splice-site variants in CTNNB1 and AMPD2 were validated by in vitro functional experiments. Interpretation: The high rate of detecting causative genetic variants (52.9%) suggests that patients diagnosed with cerebral palsy in full-term births without specific MRI findings may include genetic diseases masquerading as cerebral palsy.
Bach2 promotes B cell receptor–induced proliferation of b lymphocytes and represses cyclin-dependent kinase inhibitors Peer-reviewed

Yuichi Miura, Mizuho Morooka, Nicolas Sax, Rahul Roychoudhuri, Ari Itoh-Nakadai, Andrey Brydun, Ryo Funayama, Keiko Nakayama, Susumu Satomi, Mitsuyo Matsumoto, Kazuhiko Igarashi, Akihiko Muto

Journal of Immunology　200　(8)　2882-2893　2018/04/15
Publisher: American Association of Immunologists
DOI： 10.4049/jimmunol.1601863 　

ISSN： 1550-6606 0022-1767
Zinc finger-IRF composite elements bound by Ikaros/IRF4 complexes function as gene repression in plasma cell. Peer-reviewed

Ochiai K, Kondo H, Okamura Y, Shima H, Kurokochi Y, Kimura K, Funayama R, Nagashima T, Nakayama K, Yui K, Kinoshita K, Igarashi K

Blood advances　2　(8)　883-894　2018/04

DOI： 10.1182/bloodadvances.2017010413 　

ISSN： 2473-9529
Transforming growth factor β-induced proliferative arrest mediated by TRIM26- dependent TAF7 degradation and its antagonism by MYC Peer-reviewed

Tadashi Nakagawa, Masaki Hosogane, Makiko Nakagawa, Akane Morohoshi, Ryo Funayama, Keiko Nakayama

Molecular and Cellular Biology　38　(5)　pii: e00449-17　2018/03/01
Publisher: American Society for Microbiology
DOI： 10.1128/MCB.00449-17 　

ISSN： 1098-5549 0270-7306
Glucose deprivation induces primary cilium formation through mTORC1 inactivation Peer-reviewed

Kengo Takahashi, Tomoaki Nagai, Shuhei Chiba, Keiko Nakayama, Kensaku Mizuno

Journal of Cell Science　131　(1)　pii: jcs208769　2018/01/01
Publisher: Company of Biologists Ltd
DOI： 10.1242/jcs.208769 　

ISSN： 1477-9137 0021-9533
Inferring evolutionary responses of Anolis carolinensis introduced into the Ogasawara archipelago using whole genome sequence data Peer-reviewed

Satoshi Tamate, Watal M. Iwasaki, Kenneth L. Krysko, Brian J. Camposano, Hideaki Mori, Ryo Funayama, Keiko Nakayama, Takashi Makino, Masakado Kawata

SCIENTIFIC REPORTS　7　(1)　18008　2017/12

DOI： 10.1038/s41598-017-17852-7 　

ISSN： 2045-2322
NOTCH2 Hajdu-Cheney Mutations Escape SCFFBW7-Dependent Proteolysis to Promote Osteoporosis. International-journal Peer-reviewed

Hidefumi Fukushima, Kouhei Shimizu, Asami Watahiki, Seira Hoshikawa, Tomoki Kosho, Daiju Oba, Seiji Sakano, Makiko Arakaki, Aya Yamada, Katsuyuki Nagashima, Koji Okabe, Satoshi Fukumoto, Eijiro Jimi, Anna Bigas, Keiichi I Nakayama, Keiko Nakayama, Yoko Aoki, Wenyi Wei, Hiroyuki Inuzuka

Molecular cell　68　(4)　645-658　2017/11/16

DOI： 10.1016/j.molcel.2017.10.018 　

More details Close

Hajdu-Cheney syndrome (HCS), a rare autosomal disorder caused by heterozygous mutations in NOTCH2, is clinically characterized by acro-osteolysis, severe osteoporosis, short stature, neurological symptoms, cardiovascular defects, and polycystic kidneys. Recent studies identified that aberrant NOTCH2 signaling and consequent osteoclast hyperactivity are closely associated with the bone-related disorder pathogenesis, but the exact molecular mechanisms remain unclear. Here, we demonstrate that sustained osteoclast activity is largely due to accumulation of NOTCH2 carrying a truncated C terminus that escapes FBW7-mediated ubiquitination and degradation. Mice with osteoclast-specific Fbw7 ablation revealed osteoporotic phenotypes reminiscent of HCS, due to elevated Notch2 signaling. Importantly, administration of Notch inhibitors in Fbw7 conditional knockout mice alleviated progressive bone resorption. These findings highlight the molecular basis of HCS pathogenesis and provide clinical insights into potential targeted therapeutic strategies for skeletal disorders associated with the aberrant FBW7/NOTCH2 pathway as observed in patients with HCS.
Inflammatory responses induce an identity crisis of alveolar macrophages, leading to pulmonary alveolar proteinosis Peer-reviewed

Risa Ebina-Shibuya, Mitsuyo Matsumoto, Makoto Kuwahara, Kyoung-Jin Jang, Manabu Sugai, Yoshiaki Ito, Ryo Funayama, Keiko Nakayama, Yuki Sato, Naoto Ishii, Yasunobu Okamura, Kengo Kinoshita, Kohei Kometani, Tomohiro Kurosaki, Akihiko Muto, Masakazu Ichinose, Masakatsu Yamashita, Kazuhiko Igarashi

JOURNAL OF BIOLOGICAL CHEMISTRY　292　(44)　18098-18112　2017/11

DOI： 10.1074/jbc.M117.808535 　

ISSN： 0021-9258

eISSN： 1083-351X
Regulation of mitosis-meiosis transition by the ubiquitin ligase beta-TrCP in male germ cells Peer-reviewed

Tadashi Nakagawa, Teng Zhang, Ryo Kushi, Seiji Nakano, Takahiro Endo, Makiko Nakagawa, Noriko Yanagihara, David Zarkower, Keiko Nakayama

DEVELOPMENT　144　(22)　4137-4147　2017/11

DOI： 10.1242/dev.158485 　

ISSN： 0950-1991

eISSN： 1477-9129
Long-term outcome of a 26-year-old woman with West syndrome and an nuclear receptor subfamily 2 group F member 1 gene (NR2F1) mutation. International-journal Peer-reviewed

Naomi Hino-Fukuyo, Atsuo Kikuchi, Hiroyuki Yokoyama, Kazuie Iinuma, Mieko Hirose, Kazuhiro Haginoya, Tetsuya Niihori, Keiko Nakayama, Yoko Aoki, Shigeo Kure

Seizure　50　144-146　2017/08

DOI： 10.1016/j.seizure.2017.06.018 　

More details Close

Long-term outcome of West syndrome with a NR2F1 mutation.
Exome sequencing deciphers a germline MET mutation in familial epidermal growth factor receptor-mutant lung cancer. International-journal Peer-reviewed

Naoki Tode, Toshiaki Kikuchi, Tomohiro Sakakibara, Taizou Hirano, Akira Inoue, Shinya Ohkouchi, Tsutomu Tamada, Tatsuma Okazaki, Akira Koarai, Hisatoshi Sugiura, Tetsuya Niihori, Yoko Aoki, Keiko Nakayama, Kunio Matsumoto, Yoichi Matsubara, Masayuki Yamamoto, Akira Watanabe, Toshihiro Nukiwa, Masakazu Ichinose

Cancer science　108　(6)　1263-1270　2017/06

DOI： 10.1111/cas.13233 　

More details Close

Lung cancer accompanied by somatic activating mutations in the epidermal growth factor receptor (EGFR) gene, which is associated with a significant clinical response to the targeted therapy, is frequently found in never-smoking Asian women with adenocarcinoma. Although this implies genetic factors underlying the carcinogenesis, the etiology remains unclear. To gain insight into the pathogenic mechanisms, we sequenced the exomes in the peripheral-blood DNA from six siblings, four affected and two unaffected siblings, of a family with familial EGFR-mutant lung adenocarcinoma. We identified a heterozygous missense mutation in MET proto-oncogene, p.Asn375Lys, in all four affected siblings. Combined with somatic loss of heterozygosity for MET, the higher allele frequency in a Japanese sequencing database supports a causative role of the MET mutation in EGFR-mutant lung cancer. Functional assays showed that the mutation reduces the binding affinity of MET for its ligand, hepatocyte growth factor, and damages the subsequent cellular processes, including proliferation, clonogenicity, motility and tumorigenicity. The MET mutation was further observed to abrogate the ERBB3-mediated AKT signal transduction, which is shared downstream by EGFR. These findings provide an etiological view that the MET mutation is involved in the pathogenesis of EGFR-mutant lung cancer because it generates oncogenic stress that induces compensatory EGFR activation. The identification of MET in a family with familial EGFR-mutant lung cancer is insightful to explore the pathogenic mechanism of not only familial, but also sporadic EGFR-mutant lung cancer by underscoring MET-related signaling molecules.
Hippo Signaling Suppresses Cell Ploidy and Tumorigenesis through Skp2 Peer-reviewed

Shihao Zhang, Qinghua Chen, Qingxu Liu, Yuxi Li, Xiufeng Sun, Lixin Hong, Suyuan Ji, Chengyan Liu, Jing Geng, Weiji Zhang, Zhonglei Lu, Zhen-Yu Yin, Yuanyuan Zeng, Kwang-Huei Lin, Qiao Wu, Qiyuan Li, Keiko Nakayama, Keiich I. Nakayama, Xianming Deng, Randy L. Johnson, Liang Zhu, Daming Gao, Lanfen Chen, Dawang Zhou

CANCER CELL　31　(5)　669-+　2017/05

DOI： 10.1016/j.ccell.2017.04.004 　

ISSN： 1535-6108

eISSN： 1878-3686
Regulatory signatures of liver regeneration distilled by integrative analysis of mRNA, histone methylation, and proteomics Peer-reviewed

Yoshihiro Sato, Yasutake Katoh, Mitsuyo Matsumoto, Masaki Sato, Masayuki Ebina, Ari Itoh-Nakadai, Ryo Funayama, Keiko Nakayama, Michiaki Unno, Kazuhiko Igarashi

JOURNAL OF BIOLOGICAL CHEMISTRY　292　(19)　8019-+　2017/05

DOI： 10.1074/jbc.M116.774547 　

ISSN： 0021-9258

eISSN： 1083-351X
Dramatic response after functional hemispherectomy in a patient with epileptic encephalopathy carrying a de novo COL4A1 mutation. International-journal Peer-reviewed

Naomi Hino-Fukuyo, Atsuo Kikuchi, Masaki Iwasaki, Yuko Sato, Yuki Kubota, Tomoko Kobayashi, Tojo Nakayama, Kazuhiro Haginoya, Natsuko Arai-Ichinoi, Tetsuya Niihori, Ryo Sato, Tasuku Suzuki, Hiroki Kudo, Ryo Funayama, Keiko Nakayama, Yoko Aoki, Shigeo Kure

Brain & development　39　(4)　337-340　2017/04

DOI： 10.1016/j.braindev.2016.11.006 　

More details Close

We describe the first case of a successful functional hemispherectomy in a patient with epileptic encephalopathy and a de novo collagen type IV alpha 1 (COL4A1) mutation. A 4-year-old girl was COL4A1 mutation-positive and suffered from drug-resistant epilepsy, hemiplegia, and developmental delay. Magnetic resonance imaging detected no porencephaly, and she had no cataract or renal abnormality. Following a presurgical evaluation for epilepsy, she underwent a functional hemispherectomy. She has been seizure free with no intracranial hemorrhage or other perioperative complications. Patients with a COL4A1 mutation have an increased risk for intracranial hemorrhage because of disrupted integrity in the vascular basement membrane due to the mutation. After weighing the risks and benefits to these patients, epilepsy surgery may not be absolutely contraindicated. Furthermore, pediatric neurologists should be aware of an undiagnosed COL4A1 mutation when a patient presents with an unexplained neurological phenotype, such as mild hemiparesis, even in the absence of porencephaly.
Protein-arginine deiminase 2 suppresses proliferation of colon cancer cells through protein citrullination Peer-reviewed

Ryo Funayama, Hajime Taniguchi, Masamichi Mizuma, Fumiyoshi Fujishima, Minoru Kobayashi, Shinobu Ohnuma, Michiaki Unno, Keiko Nakayama

CANCER SCIENCE　108　(4)　713-718　2017/04

DOI： 10.1111/cas.13179 　

ISSN： 1349-7006
Geminin is an indispensable inhibitor of Cdt1 in mouse embryonic stem cells Peer-reviewed

Masaki Hosogane, Lena Bosu, Emiko Fukumoto, Hidetoshi Yamada, Soichiro Sato, Keiko Nakayama

GENES TO CELLS　22　(4)　360-375　2017/04

DOI： 10.1111/gtc.12482 　

ISSN： 1356-9597

eISSN： 1365-2443
A large-scale targeted proteomics assay resource based on an in vitro human proteome Peer-reviewed

Masaki Matsumoto, Fumiko Matsuzaki, Kiyotaka Oshikawa, Naoki Goshima, Masatoshi Mori, Yoshifumi Kawamura, Koji Ogawa, Eriko Fukuda, Hirokazu Nakatsumi, Tohru Natsume, Kazuhiko Fukui, Katsuhisa Horimoto, Takeshi Nagashima, Ryo Funayama, Keiko Nakayama, Keiichi I. Nakayama

NATURE METHODS　14　(3)　251-+　2017/03

DOI： 10.1038/NMETH.4116 　

ISSN： 1548-7091

eISSN： 1548-7105
A Bach2-Cebp Gene Regulatory Network for the Commitment of Multipotent Hematopoietic Progenitors Peer-reviewed

Ari Itoh-Nakadai, Mitsuyo Matsumoto, Hiroki Kato, Junichi Sasaki, Yukihiro Uehara, Yuki Sato, Risa Ebina-Shibuya, Mizuho Morooka, Ryo Funayama, Keiko Nakayama, Kyoko Ochiai, Akihiko Muto, Kazuhiko Igarashi

CELL REPORTS　18　(10)　2401-2414　2017/03

DOI： 10.1016/j.celrep.2017.02.029 　

ISSN： 2211-1247
A Homeostatic Shift Facilitates Endoplasmic Reticulum Proteostasis through Transcriptional Integration of Proteostatic Stress Response Pathways Peer-reviewed

Liam Baird, Tadayuki Tsujita, Eri H. Kobayashi, Ryo Funayama, Takeshi Nagashima, Keiko Nakayama, Masayuki Yamamoto

MOLECULAR AND CELLULAR BIOLOGY　37　(4)　E00651-U199　2017/02

DOI： 10.1128/MCB.00439.16 　

ISSN： 0270-7306

eISSN： 1098-5549
Ubiquitylation of Ku80 by RNF126 Promotes Completion of Nonhomologous End Joining-Mediated DNA Repair Peer-reviewed

Noriko Ishida, Tadashi Nakagawa, Shun-Ichiro Iemura, Akira Yasui, Hiroki Shima, Yasutake Katoh, Yuko Nagasawa, Toru Natsume, Kazuhiko Igarashi, Keiko Nakayama

MOLECULAR AND CELLULAR BIOLOGY　37　(4)　e00347-16　2017/02

DOI： 10.1128/MCB.00347-16 　

ISSN： 0270-7306

eISSN： 1098-5549
The aryl hydrocarbon receptor AhR links atopic dermatitis and air pollution via induction of the neurotrophic factor artemin Peer-reviewed

Takanori Hidaka, Eisaku Ogawa, Eri H. Kobayashi, Takafumi Suzuki, Ryo Funayama, Takeshi Nagashima, Taku Fujimura, Setsuya Aiba, Keiko Nakayama, Ryuhei Okuyama, Masayuki Yamamoto

NATURE IMMUNOLOGY　18　(1)　64-73　2017/01

DOI： 10.1038/ni.3614 　

ISSN： 1529-2908

eISSN： 1529-2916
The SCF beta-TRCP E3 ubiquitin ligase complex targets Lipin1 for ubiquitination and degradation to promote hepatic lipogenesis Peer-reviewed

Kouhei Shimizu, Hidefumi Fukushima, Kohei Ogura, Evan C. Lien, Naoe Taira Nihira, Jinfang Zhang, Brian J. North, Ailan Guo, Katsuyuki Nagashima, Tadashi Nakagawa, Seira Hoshikawa, Asami Watahiki, Koji Okabe, Aya Yamada, Alex Toker, John M. Asara, Satoshi Fukumoto, Keiichi I. Nakayama, Keiko Nakayama, Hiroyuki Inuzuka, Wenyi Wei

SCIENCE SIGNALING　10　(460)　2017/01

DOI： 10.1126/scisignal.aah4117 　

ISSN： 1945-0877

eISSN： 1937-9145
Wnt--catenin signaling regulates ABCC3 (MRP3) transporter expression in colorectal cancer Peer-reviewed

Minoru Kobayashi, Ryo Funayama, Shinobu Ohnuma, Michiaki Unno, Keiko Nakayama

CANCER SCIENCE　107　(12)　1776-1784　2016/12

DOI： 10.1111/cas.13097 　

ISSN： 1347-9032

eISSN： 1349-7006
The double knockout of Bach1 and Bach2 in mice reveals shared compensatory mechanisms in regulating alveolar macrophage function and lung surfactant homeostasis Peer-reviewed

Risa Ebina-Shibuya, Miki Watanabe-Matsui, Mitsuyo Matsumoto, Ari Itoh-Nakadai, Ryo Funayama, Keiko Nakayama, Akihiko Muto, Kazuhiko Igarashi

JOURNAL OF BIOCHEMISTRY　160　(6)　333-344　2016/12

DOI： 10.1093/jb/mvw041 　

ISSN： 0021-924X

eISSN： 1756-2651
大腸癌で発現が抑制されるPeptidylarginine deiminase 2の機能解析 Peer-reviewed

谷口肇, 舟山亮, 小林実, 高館達之, 阿部友哉, 水間正道, 藤島史喜, 大沼忍, 内藤剛, 海野倫明, 中山啓子

日本癌学会総会記事　75回　P-1267　2016/10
Publisher: 日本癌学会
ISSN： 0546-0476
Lack of genetic variation prevents adaptation at the geographic range margin in a damselfly Peer-reviewed

Yuma Takahashi, Yoshihisa Suyama, Yu Matsuki, Ryo Funayama, Keiko Nakayama, Masakado Kawata

MOLECULAR ECOLOGY　25　(18)　4450-4460　2016/09

DOI： 10.1111/mec.13782 　

ISSN： 0962-1083

eISSN： 1365-294X
The Tohoku Medical Megabank Project: Design and Mission Peer-reviewed

Shinichi Kuriyama, Nobuo Yaegashi, Fuji Nagami, Tomohiko Arai, Yoshio Kawaguchi, Noriko Osumi, Masaki Sakaida, Yoichi Suzuki, Keiko Nakayama, Hiroaki Hashizume, Gen Tamiya, Hiroshi Kawame, Kichiya Suzuki, Atsushi Hozawa, Naoki Nakaya, Masahiro Kikuya, Hirohito Metoki, Ichiro Tsuji, Nobuo Fuse, Hideyasu Kiyomoto, Junichi Sugawara, Akito Tsuboi, Shinichi Egawa, Kiyoshi Ito, Koichi Chida, Tadashi Ishii, Hiroaki Tomita, Yasuyuki Taki, Naoko Minegishi, Naoto Ishii, Jun Yasuda, Kazuhiko Igarashi, Ritsuko Shimizu, Masao Nagasaki, Seizo Koshiba, Kengo Kinoshita, Soichi Ogishima, Takako Takai-Igarashi, Teiji Tominaga, Osamu Tanabe, Noriaki Ohuchi, Toru Shimosegawa, Shigeo Kure, Hiroshi Tanaka, Sadayoshi Ito, Jiro Hitomi, Kozo Tanno, Motoyuki Nakamura, Kuniaki Ogasawara, Seiichiro Kobayashi, Kiyomi Sakata, Mamoru Satoh, Atsushi Shimizu, Makoto Sasaki, Ryujin Endo, Kenji Sobue, Masayuki Yamamoto

JOURNAL OF EPIDEMIOLOGY　26　(9)　493-511　2016/09

DOI： 10.2188/jea.JE20150268 　

ISSN： 0917-5040
Lack of Transcription Triggers H3K27me3 Accumulation in the Gene Body Peer-reviewed

Masaki Hosogane, Ryo Funayama, Matsuyuki Shirota, Keiko Nakayama

CELL REPORTS　16　(3)　696-706　2016/07

DOI： 10.1016/j.celrep.2016.06.034 　

ISSN： 2211-1247
Human genetic variation database, a reference database of genetic variations in the Japanese population. International-journal Peer-reviewed

Koichiro Higasa, Noriko Miyake, Jun Yoshimura, Kohji Okamura, Tetsuya Niihori, Hirotomo Saitsu, Koichiro Doi, Masakazu Shimizu, Kazuhiko Nakabayashi, Yoko Aoki, Yoshinori Tsurusaki, Shinichi Morishita, Takahisa Kawaguchi, Osuke Migita, Keiko Nakayama, Mitsuko Nakashima, Jun Mitsui, Maiko Narahara, Keiko Hayashi, Ryo Funayama, Daisuke Yamaguchi, Hiroyuki Ishiura, Wen-Ya Ko, Kenichiro Hata, Takeshi Nagashima, Ryo Yamada, Yoichi Matsubara, Akihiro Umezawa, Shoji Tsuji, Naomichi Matsumoto, Fumihiko Matsuda

Journal of human genetics　61　(6)　547-53　2016/06

DOI： 10.1038/jhg.2016.12 　

More details Close

Whole-genome and -exome resequencing using next-generation sequencers is a powerful approach for identifying genomic variations that are associated with diseases. However, systematic strategies for prioritizing causative variants from many candidates to explain the disease phenotype are still far from being established, because the population-specific frequency spectrum of genetic variation has not been characterized. Here, we have collected exomic genetic variation from 1208 Japanese individuals through a collaborative effort, and aggregated the data into a prevailing catalog. In total, we identified 156 622 previously unreported variants. The allele frequencies for the majority (88.8%) were lower than 0.5% in allele frequency and predicted to be functionally deleterious. In addition, we have constructed a Japanese-specific major allele reference genome by which the number of unique mapping of the short reads in our data has increased 0.045% on average. Our results illustrate the importance of constructing an ethnicity-specific reference genome for identifying rare variants. All the collected data were centralized to a newly developed database to serve as useful resources for exploring pathogenic variations. Public access to the database is available at http://www.genome.med.kyoto-u.ac.jp/SnpDB/.
Genomewide approaches for BACH1 target genes in mouse embryonic fibroblasts showed BACH1-Pparg pathway in adipogenesis Peer-reviewed

Mitsuyo Matsumoto, Keiichi Kondo, Takuma Shiraki, Andrey Brydun, Ryo Funayama, Keiko Nakayama, Nobuo Yaegashi, Hideki Katagiri, Kazuhiko Igarashi

GENES TO CELLS　21　(6)　553-567　2016/06

DOI： 10.1111/gtc.12365 　

ISSN： 1356-9597

eISSN： 1365-2443
Nrf2 suppresses macrophage inflammatory response by blocking proinflammatory cytokine transcription. International-journal Peer-reviewed

Eri H Kobayashi, Takafumi Suzuki, Ryo Funayama, Takeshi Nagashima, Makiko Hayashi, Hiroki Sekine, Nobuyuki Tanaka, Takashi Moriguchi, Hozumi Motohashi, Keiko Nakayama, Masayuki Yamamoto

Nature communications　7　11624-11624　2016/05/23

DOI： 10.1038/ncomms11624 　

More details Close

Nrf2 (NF-E2-related factor-2) transcription factor regulates oxidative/xenobiotic stress response and also represses inflammation. However, the mechanisms how Nrf2 alleviates inflammation are still unclear. Here, we demonstrate that Nrf2 interferes with lipopolysaccharide-induced transcriptional upregulation of proinflammatory cytokines, including IL-6 and IL-1β. Chromatin immunoprecipitation (ChIP)-seq and ChIP-qPCR analyses revealed that Nrf2 binds to the proximity of these genes in macrophages and inhibits RNA Pol II recruitment. Further, we found that Nrf2-mediated inhibition is independent of the Nrf2-binding motif and reactive oxygen species level. Murine inflammatory models further demonstrated that Nrf2 interferes with IL6 induction and inflammatory phenotypes in vivo. Thus, contrary to the widely accepted view that Nrf2 suppresses inflammation through redox control, we demonstrate here that Nrf2 opposes transcriptional upregulation of proinflammatory cytokine genes. This study identifies Nrf2 as the upstream regulator of cytokine production and establishes a molecular basis for an Nrf2-mediated anti-inflammation approach.
The expression of miR-125b-5p is increased in the serum of patients with chronic hepatitis B infection and inhibits the detection of hepatitis B virus surface antigen Peer-reviewed

M. Ninomiya, Y. Kondo, O. Kimura, R. Funayama, T. Nagashima, T. Kogure, T. Morosawa, Y. Tanaka, K. Nakayama, T. Shimosegawa

JOURNAL OF VIRAL HEPATITIS　23　(5)　330-339　2016/05

DOI： 10.1111/jvh.12522 　

ISSN： 1352-0504

eISSN： 1365-2893
Mutations in MECOM, Encoding Oncoprotein EVI1, Cause Radioulnar Synostosis with Amegakaryocytic Thrombocytopenia. International-journal Peer-reviewed

Tetsuya Niihori, Meri Ouchi-Uchiyama, Yoji Sasahara, Takashi Kaneko, Yoshiko Hashii, Masahiro Irie, Atsushi Sato, Yuka Saito-Nanjo, Ryo Funayama, Takeshi Nagashima, Shin-Ichi Inoue, Keiko Nakayama, Keiichi Ozono, Shigeo Kure, Yoichi Matsubara, Masue Imaizumi, Yoko Aoki

American journal of human genetics　97　(6)　848-54　2015/12/03

DOI： 10.1016/j.ajhg.2015.10.010 　

More details Close

Radioulnar synostosis with amegakaryocytic thrombocytopenia (RUSAT) is an inherited bone marrow failure syndrome, characterized by thrombocytopenia and congenital fusion of the radius and ulna. A heterozygous HOXA11 mutation has been identified in two unrelated families as a cause of RUSAT. However, HOXA11 mutations are absent in a number of individuals with RUSAT, which suggests that other genetic loci contribute to RUSAT. In the current study, we performed whole exome sequencing in an individual with RUSAT and her healthy parents and identified a de novo missense mutation in MECOM, encoding EVI1, in the individual with RUSAT. Subsequent analysis of MECOM in two other individuals with RUSAT revealed two additional missense mutations. These three mutations were clustered within the 8(th) zinc finger motif of the C-terminal zinc finger domain of EVI1. Chromatin immunoprecipitation and qPCR assays of the regions harboring the ETS-like motif that is known as an EVI1 binding site showed a reduction in immunoprecipitated DNA for two EVI1 mutants compared with wild-type EVI1. Furthermore, reporter assays showed that MECOM mutations led to alterations in both AP-1- and TGF-β-mediated transcriptional responses. These functional assays suggest that transcriptional dysregulation by mutant EVI1 could be associated with the development of RUSAT. We report missense mutations in MECOM resulting in a Mendelian disorder that provide compelling evidence for the critical role of EVI1 in normal hematopoiesis and in the development of forelimbs and fingers in humans.
The artificial loss of Runx1 reduces the expression of quiescence-associated transcription factors in CD4(+) T lymphocytes Peer-reviewed

Won Fen Wong, Kazuyoshi Kohu, Takeshi Nagashima, Ryo Funayama, Mitsuyo Matsumoto, Elaheh Movahed, Grace Min Yi Tan, Tee Cian Yeow, Chung Yeng Looi, Mineo Kurokawa, Motomi Osato, Kazuhiko Igarashi, Keiko Nakayama, Masanobu Satake

MOLECULAR IMMUNOLOGY　68　(2)　223-233　2015/12

DOI： 10.1016/j.molimm.2015.08.012 　

ISSN： 0161-5890
S6 Kinase- and beta-TrCP2-Dependent Degradation of p19(Arf) Is Required for Cell Proliferation Peer-reviewed

Tadashi Nakagawa, Takaaki Araki, Makiko Nakagawa, Atsushi Hirao, Michiaki Unno, Keiko Nakayama

MOLECULAR AND CELLULAR BIOLOGY　35　(20)　3517-3527　2015/10

DOI： 10.1128/MCB.00343-15 　

ISSN： 0270-7306

eISSN： 1098-5549
Rho-Kinase Inhibition During Early Cardiac Development Causes Arrhythmogenic Right Ventricular Cardiomyopathy in Mice. International-journal Peer-reviewed

Alia Ellawindy, Kimio Satoh, Shinichiro Sunamura, Nobuhiro Kikuchi, Kota Suzuki, Tatsuro Minami, Shohei Ikeda, Shinichi Tanaka, Toru Shimizu, Budbazar Enkhjargal, Satoshi Miyata, Yuhto Taguchi, Tetsuya Handoh, Kenta Kobayashi, Kazuto Kobayashi, Keiko Nakayama, Masahito Miura, Hiroaki Shimokawa

Arteriosclerosis, thrombosis, and vascular biology　35　(10)　2172-84　2015/10

DOI： 10.1161/ATVBAHA.115.305872 　

More details Close

OBJECTIVE: Arrhythmogenic right ventricular cardiomyopathy (ARVC) is characterized by fibrofatty changes of the right ventricle, ventricular arrhythmias, and sudden death. Though ARVC is currently regarded as a disease of the desmosome, desmosomal gene mutations have been identified only in half of ARVC patients, suggesting the involvement of other associated mechanisms. Rho-kinase signaling is involved in the regulation of intracellular transport and organizes cytoskeletal filaments, which supports desmosomal protein complex at the myocardial cell-cell junctions. Here, we explored whether inhibition of Rho-kinase signaling is involved in the pathogenesis of ARVC. APPROACH AND RESULTS: Using 2 novel mouse models with SM22α- or αMHC-restricted overexpression of dominant-negative Rho-kinase, we show that mice with Rho-kinase inhibition in the developing heart (SM22α-restricted) spontaneously develop cardiac dilatation and dysfunction, myocardial fibrofatty changes, and ventricular arrhythmias, resulting in premature sudden death, phenotypes fulfilling the criteria of ARVC in humans. Rho-kinase inhibition in the developing heart results in the development of ARVC phenotypes in dominant-negative Rho-kinase mice through 3 mechanisms: (1) reduction of cardiac cell proliferation and ventricular wall thickness, (2) stimulation of the expression of the proadipogenic noncanonical Wnt ligand, Wnt5b, and the major adipogenic transcription factor, PPARγ (peroxisome proliferator activated receptor-γ), and inhibition of Wnt/β-catenin signaling, and (3) development of desmosomal abnormalities. These mechanisms lead to the development of cardiac dilatation and dysfunction, myocardial fibrofatty changes, and ventricular arrhythmias, ultimately resulting in sudden premature death in this ARVC mouse model. CONCLUSIONS: This study demonstrates a novel crucial role of Rho-kinase inhibition during cardiac development in the pathogenesis of ARVC in mice.
Isolated inclusion body myopathy caused by a multisystem proteinopathy-linked hnRNPA1 mutation. International-journal Peer-reviewed

Rumiko Izumi, Hitoshi Warita, Tetsuya Niihori, Toshiaki Takahashi, Maki Tateyama, Naoki Suzuki, Ayumi Nishiyama, Matsuyuki Shirota, Ryo Funayama, Keiko Nakayama, Satomi Mitsuhashi, Ichizo Nishino, Yoko Aoki, Masashi Aoki

Neurology. Genetics　1　(3)　e23-e23　2015/10

DOI： 10.1212/NXG.0000000000000023 　

More details Close

OBJECTIVE: To identify the genetic cause of isolated inclusion body myopathy (IBM) with autosomal dominant inheritance in 2 families. METHODS: Genetic investigations were performed using whole-exome and Sanger sequencing of the heterogeneous nuclear ribonucleoprotein A1 gene (hnRNPA1). The clinical and pathologic features of patients in the 2 families were evaluated with neurologic examinations, muscle imaging, and muscle biopsy. RESULTS: We identified a missense p.D314N mutation in hnRNPA1, which is also known to cause familial amyotrophic lateral sclerosis, in 2 families with IBM. The affected individuals developed muscle weakness in their 40s, which slowly progressed toward a limb-girdle pattern. Further evaluation of the affected individuals revealed no apparent motor neuron dysfunction, cognitive impairment, or bone abnormality. The muscle pathology was compatible with IBM, lacking apparent neurogenic change and inflammation. Multiple immunohistochemical analyses revealed the cytoplasmic aggregation of hnRNPA1 in close association with autophagosomes and myonuclei. Furthermore, the aberrant accumulation was characterized by coaggregation with ubiquitin, sequestome-1/p62, valosin-containing protein/p97, and a variety of RNA-binding proteins (RBPs). CONCLUSIONS: The present study expands the clinical phenotype of hnRNPA1-linked multisystem proteinopathy. Mutations in hnRNPA1, and possibly hnRNPA2B1, will be responsible for isolated IBM with a pure muscular phenotype. Although the mechanisms underlying the selective skeletal muscle involvement remain to be elucidated, the immunohistochemical results suggest a broad sequestration of RBPs by the mutated hnRNPA1.
Role of the Atg9a gene in intrauterine growth and survival of fetal mice Peer-reviewed

Takashi Kojima, Takahiro Yamada, Rina Akaishi, Itsuko Furuta, Tatsuya Saitoh, Kazuhiko Nakabayashi, Keiichi I. Nakayama, Keiko Nakayama, Shizuo Akira, Hisanori Minakami

REPRODUCTIVE BIOLOGY　15　(3)　131-138　2015/09

DOI： 10.1016/j.repbio.2015.05.001 　

ISSN： 1642-431X
Protein monoubiquitylation: targets and diverse functions Peer-reviewed

Tadashi Nakagawa, Keiko Nakayama

GENES TO CELLS　20　(7)　543-562　2015/07

DOI： 10.1111/gtc.12250 　

ISSN： 1356-9597

eISSN： 1365-2443
Genomic analysis identifies candidate pathogenic variants in 9 of 18 patients with unexplained West syndrome. International-journal Peer-reviewed

Naomi Hino-Fukuyo, Atsuo Kikuchi, Natsuko Arai-Ichinoi, Tetsuya Niihori, Ryo Sato, Tasuku Suzuki, Hiroki Kudo, Yuko Sato, Tojo Nakayama, Yosuke Kakisaka, Yuki Kubota, Tomoko Kobayashi, Ryo Funayama, Keiko Nakayama, Mitsugu Uematsu, Yoko Aoki, Kazuhiro Haginoya, Shigeo Kure

Human genetics　134　(6)　649-58　2015/06

DOI： 10.1007/s00439-015-1553-6 　

More details Close

West syndrome, which is narrowly defined as infantile spasms that occur in clusters and hypsarrhythmia on EEG, is the most common early-onset epileptic encephalopathy (EOEE). Patients with West syndrome may have clear etiologies, including perinatal events, infections, gross chromosomal abnormalities, or cases followed by other EOEEs. However, the genetic etiology of most cases of West syndrome remains unexplained. DNA from 18 patients with unexplained West syndrome was subjected to microarray-based comparative genomic hybridization (array CGH), followed by trio-based whole-exome sequencing in 14 unsolved families. We identified candidate pathogenic variants in 50% of the patients (n = 9/18). The array CGH revealed candidate pathogenic copy number variations in four cases (22%, 4/18), including an Xq28 duplication, a 16p11.2 deletion, a 16p13.1 deletion and a 19p13.2 deletion disrupting CACNA1A. Whole-exome sequencing identified candidate mutations in known epilepsy genes in five cases (36%, 5/14). Three candidate de novo mutations were identified in three cases, with two mutations occurring in two new candidate genes (NR2F1 and CACNA2D1) (21%, 3/14). Hemizygous candidate mutations in ALG13 and BRWD3 were identified in the other two cases (14%, 2/14). Evaluating a panel of 67 known EOEE genes failed to identify significant mutations. Despite the heterogeneity of unexplained West syndrome, the combination of array CGH and whole-exome sequencing is an effective means of evaluating the genetic background in unexplained West syndrome. We provide additional evidence for NR2F1 as a causative gene and for CACNA2D1 and BRWD3 as candidate genes for West syndrome.
Mutations in PIGL in a patient with Mabry syndrome. International-journal Peer-reviewed

Ikuma Fujiwara, Yoshiko Murakami, Tetsuya Niihori, Junko Kanno, Akiko Hakoda, Osamu Sakamoto, Nobuhiko Okamoto, Ryo Funayama, Takeshi Nagashima, Keiko Nakayama, Taroh Kinoshita, Shigeo Kure, Yoichi Matsubara, Yoko Aoki

American journal of medical genetics. Part A　167A　(4)　777-85　2015/04

DOI： 10.1002/ajmg.a.36987 　

More details Close

Mabry syndrome, hyperphosphatasia mental retardation syndrome (HPMRS), is an autosomal recessive disease characterized by increased serum levels of alkaline phosphatase (ALP), severe developmental delay, intellectual disability, and seizures. Recent studies have revealed mutations in PIGV, PIGW, PIGO, PGAP2, and PGAP3 (genes that encode molecules of the glycosylphosphatidylinositol (GPI)-anchor biosynthesis pathway) in patients with HPMRS. We performed whole-exome sequencing of a patient with severe intellectual disability, distinctive facial appearance, fragile nails, and persistent increased serum levels of ALP. The result revealed a compound heterozygote with a 13-bp deletion in exon 1 (c.36_48del) and a two-base deletion in exon 2 (c.254_255del) in phosphatidylinositol glycan anchor, class L (PIGL) that caused frameshifts resulting in premature terminations. The 13-bp deletion was inherited from the father, and the two-base deletion was inherited from the mother. Expressing c.36_48del or c.254_255del cDNA with an HA-tag at the C- or N-terminus in PIGL-deficient CHO cells only partially restored the surface expression of GPI-anchored proteins (GPI-APs). Nonsynonymous changes or frameshift mutations in PIGL have been identified in patients with CHIME syndrome, a rare autosomal recessive disorder characterized by colobomas, congenital heart defects, early onset migratory ichthyosiform dermatosis, intellectual disability, and ear abnormalities. Our patient did not have colobomas, congenital heart defects, or early onset migratory ichthyosiform dermatosis and hence was diagnosed with HPMRS, and not CHIME syndrome. These results suggest that frameshift mutations that result in premature termination in PIGL cause a phenotype that is consistent with HPMRS.
Phosphorylation of p27Kip1 on serine 10 is required for its binding to CRM1 and nuclear export. International-journal

Noriko Ishida, Taichi Hara, Takumi Kamura, Minoru Yoshida, Keiko Nakayama, Keiichi I Nakayama

The Journal of biological chemistry　290　(11)　6754-6754　2015/03/13

DOI： 10.1074/jbc.A114.100762 　
CRL4(VprBP) E3 Ligase Promotes Monoubiquitylation and Chromatin Binding of TET Dioxygenases Peer-reviewed

Tadashi Nakagawa, Lei Lv, Makiko Nakagawa, Yanbao Yu, Chao Yu, Ana C. D'Alessio, Keiko Nakayama, Heng-Yu Fan, Xian Chen, Yue Xiong

MOLECULAR CELL　57　(2)　247-260　2015/01

DOI： 10.1016/j.molcel.2014.12.002 　

ISSN： 1097-2765

eISSN： 1097-4164
GNE myopathy associated with congenital thrombocytopenia: a report of two siblings. International-journal Peer-reviewed

Rumiko Izumi, Tetsuya Niihori, Naoki Suzuki, Yoji Sasahara, Takeshi Rikiishi, Ayumi Nishiyama, Shuhei Nishiyama, Kaoru Endo, Masaaki Kato, Hitoshi Warita, Hidehiko Konno, Toshiaki Takahashi, Maki Tateyama, Takeshi Nagashima, Ryo Funayama, Keiko Nakayama, Shigeo Kure, Yoichi Matsubara, Yoko Aoki, Masashi Aoki

Neuromuscular disorders : NMD　24　(12)　1068-72　2014/12

DOI： 10.1016/j.nmd.2014.07.008 　

More details Close

GNE myopathy is an autosomal recessive muscular disorder caused by mutations in the gene encoding the key enzyme in sialic acid biosynthesis, UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase (GNE/MNK). Here, we report two siblings with myopathy with rimmed vacuoles and congenital thrombocytopenia who harbored two compound heterozygous GNE mutations, p.V603L and p.G739S. Thrombocytopenia, which is characterized by shortened platelet lifetime rather than ineffective thrombopoiesis, has been observed since infancy. We performed exome sequencing and array CGH to identify the underlying genetic etiology of thrombocytopenia. No pathogenic variants were detected among the known causative genes of recessively inherited thrombocytopenia; yet, candidate variants in two genes that followed an autosomal recessive mode of inheritance, including previously identified GNE mutations, were detected. Alternatively, it is possible that the decreased activity of GNE/MNK itself, which would lead to decreased sialic content in platelets, is associated with thrombocytopenia in these patients. Further investigations are required to clarify the association between GNE myopathy and the pathogenesis of thrombocytopenia.
胎生期ストレスが胎児のエピゲノム変化および精神行動に及ぼす影響の特定 Peer-reviewed

兪志前, 舟山亮, 植野和子, 成相直樹, 小島要, 小野千晶, 笠原好之, 長崎正朗, 中山啓子, 富田博秋

日本臨床精神神経薬理学会・日本神経精神薬理学会合同年会プログラム・抄録集　24回・44回　204-204　2014/11
Publisher: 日本臨床精神神経薬理学会・日本神経精神薬理学会
PEX14欠損マウスにおける大脳皮質の形態とリン脂質の解析

阿部雄一, 伊藤竜太, 中山啓子, 丸谷寿裕, 本庄雅則, 藤谷昌司, 山下俊英, 中山敬一, 藤木幸夫

日本生化学会大会プログラム・講演要旨集　87回　[3T14a-05]　2014/10
Publisher: (公社)日本生化学会
Identification of acquired mutations by whole-genome sequencing in GATA-2 deficiency evolving into myelodysplasia and acute leukemia Peer-reviewed

Tohru Fujiwara, Noriko Fukuhara, Ryo Funayama, Naoki Nariai, Mayumi Kamata, Takeshi Nagashima, Kaname Kojima, Yasushi Onishi, Yoji Sasahara, Kenichi Ishizawa, Masao Nagasaki, Keiko Nakayama, Hideo Harigae

ANNALS OF HEMATOLOGY　93　(9)　1515-1522　2014/09

DOI： 10.1007/s00277-014-2090-4 　

ISSN： 0939-5555

eISSN： 1432-0584
HCV Infection Enhances Th17 Commitment, Which Could Affect the Pathogenesis of Autoimmune Diseases Peer-reviewed

Yasuteru Kondo, Masashi Ninomiya, Osamu Kimura, Keigo Machida, Ryo Funayama, Takeshi Nagashima, Koju Kobayashi, Eiji Kakazu, Takanobu Kato, Keiko Nakayama, Michael M. C. Lai, Tooru Shimosegawa

PLOS ONE　9　(6)　e98521-e98521　2014/06

DOI： 10.1371/journal.pone.0098521 　

ISSN： 1932-6203
Skp2 suppresses apoptosis in Rb1-deficient tumours by limiting E2F1 activity Peer-reviewed

Zhonglei Lu, Frederick Bauzon, Hao Fu, Jinhua Cui, Hongling Zhao, Keiko Nakayama, Keiich I. Nakayama, Liang Zhu

NATURE COMMUNICATIONS　5　3463-3463　2014/03

DOI： 10.1038/ncomms4463 　

ISSN： 2041-1723
p27 is regulated independently of Skp2 in the absence of Cdk2 Peer-reviewed

Shuhei Kotoshiba, Lakshmi Gopinathan, Elisabeth Pfeiffenberger, Anisa Rahim, Leah A. Vardy, Keiko Nakayama, Keiichi I. Nakayama, Philipp Kaldis

BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR CELL RESEARCH　1843　(2)　436-445　2014/02

DOI： 10.1016/j.bbamcr.2013.11.005 　

ISSN： 0167-4889

eISSN： 0006-3002
RNA sequencing-based identification of aberrant imprinting in cloned mice Peer-reviewed

Hiroaki Okae, Shogo Matoba, Takeshi Nagashima, Eiji Mizutani, Kimiko Inoue, Narumi Ogonuki, Hatsune Chiba, Ryo Funayama, Satoshi Tanaka, Nobuo Yaegashi, Keiko Nakayama, Hiroyuki Sasaki, Atsuo Ogura, Takahiro Arima

HUMAN MOLECULAR GENETICS　23　(4)　992-1001　2014/02

DOI： 10.1093/hmg/ddt495 　

ISSN： 0964-6906

eISSN： 1460-2083
TBX1 mutation identified by exome sequencing in a Japanese family with 22q11.2 deletion syndrome-like craniofacial features and hypocalcemia. International-journal Peer-reviewed

Tsutomu Ogata, Tetsuya Niihori, Noriko Tanaka, Masahiko Kawai, Takeshi Nagashima, Ryo Funayama, Keiko Nakayama, Shinichi Nakashima, Fumiko Kato, Maki Fukami, Yoko Aoki, Yoichi Matsubara

PloS one　9　(3)　e91598-e91598　2014

DOI： 10.1371/journal.pone.0091598 　

More details Close

BACKGROUND: Although TBX1 mutations have been identified in patients with 22q11.2 deletion syndrome (22q11.2DS)-like phenotypes including characteristic craniofacial features, cardiovascular anomalies, hypoparathyroidism, and thymic hypoplasia, the frequency of TBX1 mutations remains rare in deletion-negative patients. Thus, it would be reasonable to perform a comprehensive genetic analysis in deletion-negative patients with 22q11.2DS-like phenotypes. METHODOLOGY/PRINCIPAL FINDINGS: We studied three subjects with craniofacial features and hypocalcemia (group 1), two subjects with craniofacial features alone (group 2), and three subjects with normal phenotype within a single Japanese family. Fluorescence in situ hybridization analysis excluded chromosome 22q11.2 deletion, and genomewide array comparative genomic hybridization analysis revealed no copy number change specific to group 1 or groups 1+2. However, exome sequencing identified a heterozygous TBX1 frameshift mutation (c.1253delA, p.Y418fsX459) specific to groups 1+2, as well as six missense variants and two in-frame microdeletions specific to groups 1+2 and two missense variants specific to group 1. The TBX1 mutation resided at exon 9C and was predicted to produce a non-functional truncated protein missing the nuclear localization signal and most of the transactivation domain. CONCLUSIONS/SIGNIFICANCE: Clinical features in groups 1+2 are well explained by the TBX1 mutation, while the clinical effects of the remaining variants are largely unknown. Thus, the results exemplify the usefulness of exome sequencing in the identification of disease-causing mutations in familial disorders. Furthermore, the results, in conjunction with the previous data, imply that TBX1 isoform C is the biologically essential variant and that TBX1 mutations are associated with a wide phenotypic spectrum, including most of 22q11.2DS phenotypes.
Skp2 Deletion Unmasks a p27 Safeguard that Blocks Tumorigenesis in the Absence of pRb and p53 Tumor Suppressors Peer-reviewed

Hongling Zhao, Frederick Bauzon, Hao Fu, Zhonglei Lu, Jinhua Cui, Keiko Nakayama, Keiich I. Nakayama, Joseph Locker, Liang Zhu

Cancer Cell　24　(5)　645-659　2013/11/11

DOI： 10.1016/j.ccr.2013.09.021 　

ISSN： 1535-6108 1878-3686
Lymphotropic HCV infection enhances Th17 commitment, which could affect the pathogenesis of autoimmune and cryoglobuline-related diseases Peer-reviewed

Kondo Yasuteru, Ninomiya Masashi, Kimura Osamu, Machida Keigo, Funayama Ryo, Nagashima Takeshi, Kobayashi Koju, Kakazu Eiji, Kogure Takayuki, Kato Takanobu, Nakayama Keiko, Shimosegawa Tooru

HEPATOLOGY　58　1187A-1188A　2013/10

ISSN： 0270-9139
Loss of Protein Kinase C-δ Protects against LPS-Induced Osteolysis Owing to an Intrinsic Defect in Osteoclastic Bone Resorption Peer-reviewed

Ee Cheng Khor, Tamara Abel, Jennifer Tickner, Shek Man Chim, Cathy Wang, Taksum Cheng, Benjamin Ng, Pei Ying Ng, Dian Astari Teguh, Jacob Kenny, Xiaohong Yang, Honghui Chen, Keiichi I. Nakayama, Keiko Nakayama, Nathan Pavlos, Ming H. Zheng, Jiake Xu

PLoS ONE　8　(8)　e70815-e70815　2013/08/08

DOI： 10.1371/journal.pone.0070815 　

ISSN： 1932-6203
Ras-Induced Changes in H3K27me3 Occur after Those in Transcriptional Activity Peer-reviewed

Masaki Hosogane, Ryo Funayama, Yuichiro Nishida, Takeshi Nagashima, Keiko Nakayama

PLOS GENETICS　9　(8)　e1003698-e1003698　2013/08

DOI： 10.1371/journal.pgen.1003698 　

ISSN： 1553-7404
Gain-of-function mutations in RIT1 cause Noonan syndrome, a RAS/MAPK pathway syndrome. International-journal Peer-reviewed

Yoko Aoki, Tetsuya Niihori, Toshihiro Banjo, Nobuhiko Okamoto, Seiji Mizuno, Kenji Kurosawa, Tsutomu Ogata, Fumio Takada, Michihiro Yano, Toru Ando, Tadataka Hoshika, Christopher Barnett, Hirofumi Ohashi, Hiroshi Kawame, Tomonobu Hasegawa, Takahiro Okutani, Tatsuo Nagashima, Satoshi Hasegawa, Ryo Funayama, Takeshi Nagashima, Keiko Nakayama, Shin-Ichi Inoue, Yusuke Watanabe, Toshihiko Ogura, Yoichi Matsubara

American journal of human genetics　93　(1)　173-80　2013/07/11

DOI： 10.1016/j.ajhg.2013.05.021 　

More details Close

RAS GTPases mediate a wide variety of cellular functions, including cell proliferation, survival, and differentiation. Recent studies have revealed that germline mutations and mosaicism for classical RAS mutations, including those in HRAS, KRAS, and NRAS, cause a wide spectrum of genetic disorders. These include Noonan syndrome and related disorders (RAS/mitogen-activated protein kinase [RAS/MAPK] pathway syndromes, or RASopathies), nevus sebaceous, and Schimmelpenning syndrome. In the present study, we identified a total of nine missense, nonsynonymous mutations in RIT1, encoding a member of the RAS subfamily, in 17 of 180 individuals (9%) with Noonan syndrome or a related condition but with no detectable mutations in known Noonan-related genes. Clinical manifestations in the RIT1-mutation-positive individuals are consistent with those of Noonan syndrome, which is characterized by distinctive facial features, short stature, and congenital heart defects. Seventy percent of mutation-positive individuals presented with hypertrophic cardiomyopathy; this frequency is high relative to the overall 20% incidence in individuals with Noonan syndrome. Luciferase assays in NIH 3T3 cells showed that five RIT1 alterations identified in children with Noonan syndrome enhanced ELK1 transactivation. The introduction of mRNAs of mutant RIT1 into 1-cell-stage zebrafish embryos was found to result in a significant increase of embryos with craniofacial abnormalities, incomplete looping, a hypoplastic chamber in the heart, and an elongated yolk sac. These results demonstrate that gain-of-function mutations in RIT1 cause Noonan syndrome and show a similar biological effect to mutations in other RASopathy-related genes.
Bach1 is critical for the transformation of mouse embryonic fibroblasts by Ras(V12) and maintains ERK signaling Peer-reviewed

A. Nakanome, A. Brydun, M. Matsumoto, K. Ota, R. Funayama, K. Nakayama, M. Ono, K. Shiga, T. Kobayashi, K. Igarashi

ONCOGENE　32　(27)　3231-3245　2013/07

DOI： 10.1038/onc.2012.336 　

ISSN： 0950-9232
Y chromosome-linked B and NK cell deficiency in mice Peer-reviewed

Shu-Lan Sun, Satoshi Horino, Ari Itoh-Nakadai, Takeshi Kawabe, Atsuko Asao, Takeshi Takahashi, Takanori So, Ryo Funayama, Motonari Kondo, Hirotomo Saitsu, Naomichi Matsumoto, Keiko Nakayama, Naoto Ishii

Journal of Immunology　190　(12)　6209-6220　2013/06/15

DOI： 10.4049/jimmunol.1300303 　

ISSN： 0022-1767 1550-6606
Distinct MicroRNAs Expression Profile in Primary Biliary Cirrhosis and Evaluation of miR 505-3p and miR197-3p as Novel Biomarkers Peer-reviewed

Masashi Ninomiya, Yasuteru Kondo, Ryo Funayama, Takeshi Nagashima, Takayuki Kogure, Eiji Kakazu, Osamu Kimura, Yoshiyuki Ueno, Keiko Nakayama, Tooru Shimosegawa

PLOS ONE　8　(6)　e66086-e66086　2013/06

DOI： 10.1371/journal.pone.0066086 　

ISSN： 1932-6203
Exome sequencing identifies a novel TTN mutation in a family with hereditary myopathy with early respiratory failure. International-journal Peer-reviewed

Rumiko Izumi, Tetsuya Niihori, Yoko Aoki, Naoki Suzuki, Masaaki Kato, Hitoshi Warita, Toshiaki Takahashi, Maki Tateyama, Takeshi Nagashima, Ryo Funayama, Koji Abe, Keiko Nakayama, Masashi Aoki, Yoichi Matsubara

Journal of human genetics　58　(5)　259-66　2013/05

DOI： 10.1038/jhg.2013.9 　

More details Close

Myofibrillar myopathy (MFM) is a group of chronic muscular disorders that show the focal dissolution of myofibrils and accumulation of degradation products. The major genetic basis of MFMs is unknown. In 1993, our group reported a Japanese family with dominantly inherited cytoplasmic body myopathy, which is now included in MFM, characterized by late-onset chronic progressive distal muscle weakness and early respiratory failure. In this study, we performed linkage analysis and exome sequencing on these patients and identified a novel c.90263G>T mutation in the TTN gene (NM_001256850). During the course of our study, another groups reported three mutations in TTN in patients with hereditary myopathy with early respiratory failure (HMERF, MIM #603689), which is characterized by overlapping pathologic findings with MFMs. Our patients were clinically compatible with HMERF. The mutation identified in this study and the three mutations in patients with HMERF were located on the A-band domain of titin, suggesting a strong relationship between mutations in the A-band domain of titin and HMERF. Mutation screening of TTN has been rarely carried out because of its huge size, consisting of 363 exons. It is possible that focused analysis of TTN may detect more mutations in patients with MFMs, especially in those with early respiratory failure.
原発性胆汁性肝硬変における治療前後での血清内microRNAの発現の変化

二宮匡史, 近藤泰輝, 小暮高之, 嘉数英二, 木村修, 舟山亮, 長嶋剛史, 中山啓子, 下瀬川徹

日本臨床分子医学会学術総会プログラム・抄録集　50回　68-68　2013/04
Publisher: 日本臨床分子医学会
Opposing functions of Fbxw7 in keratinocyte growth, differentiation and skin tumorigenesis mediated through negative regulation of c-Myc and Notch Peer-reviewed

Y. Ishikawa, M. Hosogane, R. Okuyama, S. Aoyama, I. Onoyama, K. I. Nakayama, K. Nakayama

ONCOGENE　32　(15)　1921-1932　2013/04

DOI： 10.1038/onc.2012.213 　

ISSN： 0950-9232
Protein Kinase C-Delta Deficiency Perturbs Bone Homeostasis by Selective Uncoupling of Cathepsin K Secretion and Ruffled Border Formation in Osteoclasts Peer-reviewed

Viviana Cremasco, Corinne E. Decker, Deborah Stumpo, Perry J. Blackshear, Keiichi I. Nakayama, Keiko Nakayama, Traian S. Lupu, Daniel B. Graham, Deborah V. Novack, Roberta Faccio

JOURNAL OF BONE AND MINERAL RESEARCH　27　(12)　2452-2463　2012/12

DOI： 10.1002/jbmr.1701 　

ISSN： 0884-0431
Nrf2-MafG heterodimers contribute globally to antioxidant and metabolic networks Peer-reviewed

Yosuke Hirotsu, Fumiki Katsuoka, Ryo Funayama, Takeshi Nagashima, Yuichiro Nishida, Keiko Nakayama, James Douglas Engel, Masayuki Yamamoto

NUCLEIC ACIDS RESEARCH　40　(20)　10228-10239　2012/11

DOI： 10.1093/nar/gks827 　

ISSN： 0305-1048

eISSN： 1362-4962
Development of mice without Cip/Kip CDK inhibitors Peer-reviewed

Yuki Tateishi, Akinobu Matsumoto, Tomoharu Kanie, Eiji Hara, Keiko Nakayama, Keiichi I. Nakayama

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS　427　(2)　285-292　2012/10

DOI： 10.1016/j.bbrc.2012.09.041 　

ISSN： 0006-291X
Acetylation-Dependent Regulation of Skp2 Function Peer-reviewed

Hiroyuki Inuzuka, Daming Gao, Lydia W. S. Finley, Wen Yang, Lixin Wan, Hidefumi Fukushima, Y. Rebecca Chin, Bo Zhai, Shavali Shaik, Alan W. Lau, Zhiwei Wang, Steven P. Gygi, Keiko Nakayama, Julie Teruya-Feldstein, Alex Toker, Marcia C. Haigis, Pier Paolo Pandolfi, Wenyi Wei

CELL　150　(1)　179-193　2012/07

DOI： 10.1016/j.cell.2012.05.038 　

ISSN： 0092-8674
SCF(Fbw7) modulates the NFkB signaling pathway by targeting NFkB2 for ubiquitination and destruction. International-journal Peer-reviewed

Hidefumi Fukushima, Akinobu Matsumoto, Hiroyuki Inuzuka, Bo Zhai, Alan W Lau, Lixin Wan, Daming Gao, Shavali Shaik, Min Yuan, Steven P Gygi, Eijiro Jimi, John M Asara, Keiko Nakayama, Keiichi I Nakayama, Wenyi Wei

Cell reports　1　(5)　434-43　2012/05/31

DOI： 10.1016/j.celrep.2012.04.002 　

eISSN： 2211-1247

More details Close

The NFkB/Rel family of proteins play critical roles in a variety of cellular processes. Thus, their physiological activation is tightly controlled. Recently, the NFkB2/p100 precursor has been characterized as the fourth IkB type of suppressor for NFkB. However, the molecular mechanism(s) underlying regulated destruction of NFkB2 remains largely unknown. Here, we report that, unlike other IkBs, ubiquitination and destruction of NFkB2 are governed by SCF(Fbw7) in a GSK3-dependent manner. In Fbw(7-/-) cells, elevated expression of NFkB2/p100 leads to a subsequent reduction in NFkB signaling pathways and elevated sensitivity to TNFa-induced cell death. Reintroducing wild-type Fbw7, but not disease-derived mutant forms of Fbw7, rescues NFkB activity. Furthermore, T cell-specific depletion of Fbw7 also leads to reduced NFkB activity and perturbed T cell differentiation. Therefore, our work identifies Fbw7 as a physiological E3 ligase controlling NFkB20s stability. It further implicates that Fbw7 might exert its tumor-suppressor function by regulating NFkB activity.
The Amelioration of Renal Damage in Skp2-Deficient Mice Canceled by p27 (Kip1) Deficiency in Skp2(-/-) p27(-/-) Mice Peer-reviewed

Sayuri Suzuki, Hirotaka Fukasawa, Taro Misaki, Akashi Togawa, Naro Ohashi, Kyoko Kitagawa, Yojiro Kotake, Ning Liu, Hiroyuki Niida, Keiko Nakayama, Keiichi I. Nakayama, Tatsuo Yamamoto, Masatoshi Kitagawa

PLOS ONE　7　(4)　e36249-e36249　2012/04

DOI： 10.1371/journal.pone.0036249 　

ISSN： 1932-6203
Use of Illumina Deep Sequencing Technology To Differentiate Hepatitis C Virus Variants Peer-reviewed

Masashi Ninomiya, Yoshiyuki Ueno, Ryo Funayama, Takeshi Nagashima, Yuichiro Nishida, Yasuteru Kondo, Jun Inoue, Eiji Kakazu, Osamu Kimura, Keiko Nakayama, Tooru Shimosegawa

JOURNAL OF CLINICAL MICROBIOLOGY　50　(3)　857-866　2012/03

DOI： 10.1128/JCM.05715-11 　

ISSN： 0095-1137
Re-investigation and RNA sequencing-based identification of genes with placenta-specific imprinted expression Peer-reviewed

Hiroaki Okae, Hitoshi Hiura, Yuichiro Nishida, Ryo Funayama, Satoshi Tanaka, Hatsune Chiba, Nobuo Yaegashi, Keiko Nakayama, Hiroyuki Sasaki, Takahiro Arima

HUMAN MOLECULAR GENETICS　21　(3)　548-558　2012/02

DOI： 10.1093/hmg/ddr488 　

ISSN： 0964-6906
Corneal Antifibrotic Switch Identified in Genetic and Pharmacological Deficiency of Vimentin Peer-reviewed

Paola Bargagna-Mohan, Riya R. Paranthan, Adel Hamza, Chang-Guo Zhan, Do-Min Lee, Kyung Bo Kim, Daniel L. Lau, Cidambi Srinivasan, Keiko Nakayama, Keiichi I. Nakayama, Harald Herrmann, Royce Mohan

JOURNAL OF BIOLOGICAL CHEMISTRY　287　(2)　989-1006　2012/01

DOI： 10.1074/jbc.M111.297150 　

ISSN： 0021-9258
Deficient p27 Phosphorylation at Serine 10 Increases Macrophage Foam Cell Formation and Aggravates Atherosclerosis Through a Proliferation-Independent Mechanism Invited Peer-reviewed

Jose J. Fuster, Herminia Gonzalez-Navarro, Angela Vinue, Pedro Molina-Sanchez, Maria J. Andres-Manzano, Keiichi I. Nakayama, Keiko Nakayama, Antonio Diez-Juan, Antonio Bernad, Cristina Rodriguez, Jose Martinez-Gonzalez, Vicente Andres

ARTERIOSCLEROSIS THROMBOSIS AND VASCULAR BIOLOGY　31　(11)　2455-U252　2011/11

DOI： 10.1161/ATVBAHA.111.235580 　

ISSN： 1079-5642
Deregulation of the p57-E2F1-p53 Axis Results in Nonobstructive Hydrocephalus and Cerebellar Malformation in Mice Invited Peer-reviewed

Akinobu Matsumoto, Etsuo Susaki, Ichiro Onoyama, Keiko Nakayama, Mikio Hoshino, Keiichi I. Nakayama

MOLECULAR AND CELLULAR BIOLOGY　31　(20)　4176-4192　2011/10

DOI： 10.1128/MCB.05370-11 　

ISSN： 0270-7306
p57(KiP2) and p27(Kip1) Cooperate to Maintain Hematopoietic Stem Cell Quiescence through Interactions with Hsc70 Invited Peer-reviewed

Peng Zou, Hiroki Yoshihara, Kentaro Hosokawa, Ikue Tai, Kaori Shinmyozu, Fujiko Tsukahara, Yoshiro Maru, Keiko Nakayama, Keiichi I. Nakayama, Toshio Suda

CELL STEM CELL　9　(3)　247-261　2011/09

DOI： 10.1016/j.stem.2011.07.003 　

ISSN： 1934-5909
p57 Is Required for Quiescence and Maintenance of Adult Hematopoietic Stem Cells Peer-reviewed

Akinobu Matsumoto, Shoichiro Takeishi, Tomoharu Kanie, Etsuo Susaki, Ichiro Onoyama, Yuki Tateishi, Keiko Nakayama, Keiichi I. Nakayama

CELL STEM CELL　9　(3)　262-271　2011/09

DOI： 10.1016/j.stem.2011.06.014 　

ISSN： 1934-5909
Impaired ovarian development and reduced fertility in female mice deficient in Skp2 Peer-reviewed

Abbas Fotovati, Samah Abu-Ali, Keiko Nakayama, Keiichi I. Nakayama

JOURNAL OF ANATOMY　218　(6)　668-677　2011/06

DOI： 10.1111/j.1469-7580.2011.01370.x 　

ISSN： 0021-8782
Fbxw7β resides in the endoplasmic reticulum membrane and protects cells from oxidative stress. International-journal Peer-reviewed

Akinobu Matsumoto, Yuki Tateishi, Ichiro Onoyama, Yasutaka Okita, Keiko Nakayama, Keiichi I Nakayama

Cancer science　102　(4)　749-55　2011/04

DOI： 10.1111/j.1349-7006.2011.01851.x 　

More details Close

Oxidative stress has been implicated in cancer initiation and progression. Fbxw7 (also known as Fbw7, SEL-10, hCdc4, or hAgo) is the F-box protein subunit of an Skp1-Cul1-F-box (SCF)-type ubiquitin ligase complex that plays a central role in the degradation of oncoproteins such as c-Myc, c-Jun, Notch, and cyclin E. Fbxw7 is therefore thought to function as a tumor suppressor, and indeed the Fbxw7 gene is frequently mutated in many human malignancies. The Fbxw7 gene locus encodes three protein isoforms: Fbxw7α, Fbxw7β, and Fbxw7γ. Whereas Fbxw7α and Fbxw7γ are resident in the nucleus, Fbxw7β shows a cytoplasmic distribution suggestive of localization to the endoplasmic reticulum (ER). The specific function of Fbxw7β has remained unknown, however. We now show that Fbxw7β contains a putative transmembrane domain near its NH(2) -terminus, and topological analysis revealed that Fbxw7β is inserted in the ER membrane. Fbxw7β assembled with Skp1, Cul1, and Rbx1 to form an SCF complex, although the efficiency of this process appeared lower than that for Fbxw7α or Fbxw7γ. To explore the physiological role of Fbxw7β, we generated mice specifically lacking this isoform of Fbxw7. Although these animals did not exhibit any apparent abnormalities in development, primary cultures of neurons prepared from the mutant mice were more vulnerable to oxidative stress than were those prepared from wild-type mice. Conversely, overexpression of Fbxw7β rendered cells resistant to oxidative stress, without affecting sensitivity to ER stress or other apoptosis-inducing agents. Our results thus suggest that Fbxw7β contributes to the protection of cells from oxidative stress.
Fbxw7 regulates lipid metabolism and cell fate decisions in the mouse liver. International-journal Peer-reviewed

Ichiro Onoyama, Atsushi Suzuki, Akinobu Matsumoto, Kengo Tomita, Hideki Katagiri, Yuichi Oike, Keiko Nakayama, Keiichi I Nakayama

The Journal of clinical investigation　121　(1)　342-54　2011/01

DOI： 10.1172/JCI40725 　

More details Close

E3 ubiquitin ligase complexes of the SCF type consist of ring-box 1 (Rbx1), cullin 1 (Cul1), S-phase kinase-associated protein 1 (Skp1), and a member of the F-box family of proteins. The identity of the F-box protein determines the substrate specificity of the complex. The F-box family member F-box- and WD repeat domain-containing 7 (Fbxw7; also known as Fbw7, SEL-10, hCdc4, and hAgo) targets for degradation proteins with wide-ranging functions, and uncovering its in vivo role has been difficult, because Fbxw7-/- embryos die in utero. Using two different Cre-loxP systems (Mx1-Cre and Alb-Cre), we generated mice with liver-specific null mutations of Fbxw7. Hepatic ablation of Fbxw7 resulted in hepatomegaly and steatohepatitis, with massive deposition of triglyceride, a phenotype similar to that observed in humans with nonalcoholic steatohepatitis. Both cell proliferation and the abundance of Fbxw7 substrates were increased in the Fbxw7-deficient liver. Long-term Fbxw7 deficiency resulted in marked proliferation of the biliary system and the development of hamartomas. Fbxw7 deficiency also skewed the differentiation of liver stem cells toward the cholangiocyte lineage rather than the hepatocyte lineage in vitro. This bias was corrected by additional loss of the Notch cofactor RBP-J, suggesting that Notch accumulation triggered the abnormal proliferation of the biliary system. Together, our results suggest that Fbxw7 plays key roles, regulating lipogenesis and cell proliferation and differentiation in the liver.
Localization of SMAP2 to the TGN and its Function in the Regulation of TGN Protein Transport Invited Peer-reviewed

Tomo Funaki, Shunsuke Kon, Roger E. Ronn, Yuji Henmi, Yuka Kobayashi, Toshio Watanabe, Keiko Nakayama, Kenji Tanabe, Masanobu Satake

CELL STRUCTURE AND FUNCTION　36　(1)　83-95　2011

DOI： 10.1247/csf.10022 　

ISSN： 0386-7196

eISSN： 1347-3700
A Diacylglycerol-Dependent Signaling Pathway Contributes to Regulation of Antibacterial Autophagy Peer-reviewed

Shahab Shahnazari, Wei-Lien Yen, Cheryl L. Birmingham, Jessica Shiu, Anton Namolovan, Yiyu T. Zheng, Keiko Nakayama, Daniel J. Klionsky, John H. Brumell

CELL HOST & MICROBE　8　(2)　137-146　2010/08

DOI： 10.1016/j.chom.2010.07.002 　

ISSN： 1931-3128
Nuclear IKK beta Is an Adaptor Protein for I kappa B alpha Ubiquitination and Degradation in UV-Induced NF-kappa B Activation Invited Peer-reviewed

Yoshihiro Tsuchiya, Tomoichiro Asano, Keiko Nakayama, Tomohisa Kato, Michael Karin, Hideaki Kamata

MOLECULAR CELL　39　(4)　570-582　2010/08

DOI： 10.1016/j.molcel.2010.07.030 　

ISSN： 1097-2765
Inhibition of FOXO3 Tumor Suppressor Function by beta TrCP1 through Ubiquitin-Mediated Degradation in a Tumor Mouse Model Peer-reviewed

Wen-Bin Tsai, Young Min Chung, Yiyu Zou, See-Hyoung Park, Zhaohui Xu, Keiko Nakayama, Sue-Hwa Lin, Mickey C-T. Hu

PLOS ONE　5　(7)　e11171-e11171　2010/07

DOI： 10.1371/journal.pone.0011171 　

ISSN： 1932-6203
Complex regulation of cell-cycle inhibitors by Fbxw7 in mouse embryonic fibroblasts Peer-reviewed

K. Masuda, Y. Ishikawa, I. Onoyama, M. Unno, I. M. de Alboran, K. I. Nakayama, K. Nakayama

ONCOGENE　29　(12)　1798-1809　2010/03

DOI： 10.1038/onc.2009.469 　

ISSN： 0950-9232
Skp2 is required for survival of aberrantly proliferating Rb1-deficient cells and for tumorigenesis in Rb1(+/-) mice Peer-reviewed

Hongbo Wang, Frederick Bauzon, Peng Ji, Xiaoliang Xu, Daqian Sun, Joseph Locker, Rani S. Sellers, Keiko Nakayama, Keiich I. Nakayama, David Cobrinik, Liang Zhu

NATURE GENETICS　42　(1)　83-U105　2010/01

DOI： 10.1038/ng.498 　

ISSN： 1061-4036

eISSN： 1546-1718
S-phase kinase-associated protein-2 (Skp2) promotes vascular smooth muscle cell proliferation and neointima formation in vivo Peer-reviewed

Yih-Jer Wu, Graciela B. Sala-Newby, Kuo-Tung Shu, Hung-I Yeh, Keiichi I. Nakayama, Keiko Nakayama, Andrew C. Newby, Mark Bond

JOURNAL OF VASCULAR SURGERY　50　(5)　1135-1142　2009/11

DOI： 10.1016/j.jvs.2009.07.066 　

ISSN： 0741-5214
Mechanoregulation of Proliferation Peer-reviewed

Xiaogang Jiang, Paul F. Austin, Robert A. Niederhoff, Scott R. Manson, Jacob J. Riehm, Brian L. Cook, Gina Pengue, Kanchan Chitaley, Keiko Nakayama, Keiichi I. Nakayama, Steven J. Weintraub

MOLECULAR AND CELLULAR BIOLOGY　29　(18)　5104-5114　2009/09

DOI： 10.1128/MCB.00465-09 　

ISSN： 0270-7306
Human synovial sarcoma proto-oncogene Syt is essential for early embryonic development through the regulation of cell migration Peer-reviewed

Taichi Kimura, Mieko Sakai, Kouichi Tabu, Lei Wang, Ryosuke Tsunematsu, Masumi Tsuda, Hirofumi Sawa, Kazuo Nagashima, Hiroshi Nishihara, Shigetsugu Hatakeyama, Keiko Nakayama, Marc Ladanyi, Shinya Tanaka, Keiichi I. Nakayama

LABORATORY INVESTIGATION　89　(6)　645-656　2009/06

DOI： 10.1038/labinvest.2009.25 　

ISSN： 0023-6837
Protein Kinase C delta Differentially Regulates Platelet Functional Responses Peer-reviewed

Ramya Chari, Todd Getz, Bela Nagy, Kamala Bhavaraju, Yingying Mao, Yamini Saraswathy Bynagari, Swaminathan Murugappan, Keiko Nakayama, Satya P. Kunapuli

ARTERIOSCLEROSIS THROMBOSIS AND VASCULAR BIOLOGY　29　(5)　699-705　2009/05

DOI： 10.1161/ATVBAHA.109.184010 　

ISSN： 1079-5642
Common and specific roles of the related CDK inhibitors p27 and p57 revealed by a knock-in mouse model Peer-reviewed

Etsuo Susaki, Keiko Nakayama, Lili Yamasaki, Keiichi I. Nakayama

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA　106　(13)　5192-5197　2009/03

DOI： 10.1073/pnas.0811712106 　

ISSN： 0027-8424
組織特異的発がん機構の解明 Invited

中山啓子

上原記念生命科学財団研究報告集　22　1-4　2008/12
The Role of beta-TrCP1 and beta-TrCP2 in Circadian Rhythm Generation by Mediating Degradation of Clock Protein PER2 Peer-reviewed

Kanae Ohsaki, Katsutaka Oishi, Yuko Kozono, Keiko Nakayama, Keiichi I. Nakayama, Norio Ishida

JOURNAL OF BIOCHEMISTRY　144　(5)　609-618　2008/11

DOI： 10.1093/jb/mvn112 　

ISSN： 0021-924X

eISSN： 1756-2651
A functional link between Wnt signaling and SKP2-independent p27 turnover in mammary tumors Peer-reviewed

Gustavo A. Miranda-Carboni, Susan A. Krum, Kathleen Yee, Miguel Nava, Qiming E. Deng, Shehla Pervin, Alicia Collado-Hidalgo, Zoran Galic, Jerome A. Zack, Keiko Nakayama, Keiichi I. Nakayama, Timothy F. Lane

GENES & DEVELOPMENT　22　(22)　3121-3134　2008/11

DOI： 10.1101/gad.1692808 　

ISSN： 0890-9369
Notch-dependent cell cycle arrest and apoptosis in mouse embryonic fibroblasts lacking Fbxw7 Peer-reviewed

Y. Ishikawa, I. Onoyama, K. I. Nakayama, K. Nakayama

ONCOGENE　27　(47)　6164-6174　2008/10

DOI： 10.1038/onc.2008.216 　

ISSN： 0950-9232
Skp2 regulates the antiproliferative function of the tumor suppressor RASSF1A via ubiquitin-mediated degradation at the G(1)-S transition Peer-reviewed

M. S. Song, S. J. Song, S. J. Kim, K. Nakayama, K. I. Nakayama, D-S Lim

ONCOGENE　27　(22)　3176-3185　2008/05

DOI： 10.1038/sj.onc.1210971 　

ISSN： 0950-9232
Targeting the p27 E3 ligase SCFSkp2 results in p27-and Skp2-mediated cell-cycle arrest and activation of autophagy Peer-reviewed

Qing Chen, Weilin Xie, Deborah J. Kuhn, Peter M. Voorhees, Antonia Lopez-Girona, Derek Mendy, Laura G. Corral, Veronique Plantevin Krenitsky, Weiming Xu, Laure Moutouh-de Parseval, David R. Webb, Frank Mercurio, Keiichi I. Nakayama, Keiko Nakayama, Robert Z. Orlowski

BLOOD　111　(9)　4690-4699　2008/05

DOI： 10.1182/blood-2007-09-112904 　

ISSN： 0006-4971
Fbxw7 acts as a critical fail-safe against premature loss of hematopoietic stem cells and development of T-ALL Peer-reviewed

Sahoko Matsuoka, Yuichi Oike, Ichiro Onoyama, Atsushi Iwama, Fumio Arai, Keiyo Takubo, Yoichi Mashimo, Hideyuki Oguro, Eriko Nitta, Keisuke Ito, Kana Miyamoto, Hiroki Yoshiwara, Kentaro Hosokawa, Yuka Nakamura, Yumiko Gomei, Hiroko Iwasaki, Yasuhide Hayashi, Yumi Matsuzaki, Keiko Nakayama, Yasuo Ikeda, Akira Hata, Shigeru Chiba, Keiichi I. Nakayama, Toshio Suda

GENES & DEVELOPMENT　22　(8)　986-991　2008/04

DOI： 10.1101/gad.1621808 　

ISSN： 0890-9369
【細胞内の輪廻転生タンパク質の分解機構ユビキチン、プロテアソーム、オートファジー、プロテオリシスなど分解装置の作動機構と病態生理作用】ユビキチン SCF型ユビキチンリガーゼによる細胞周期制御と発癌 Invited Peer-reviewed

中山敬一, 中山啓子

実験医学　26　(2)　166-174　2008/02
SCF beta TrCP1 activates and ubiquitylates TAp63 gamma Invited Peer-reviewed

Jayme R. Gallegos, Joel Litersky, Hunjoo Lee, Yi Sun, Keiichi Nakayama, Keiko Nakayama, Hua Lu

JOURNAL OF BIOLOGICAL CHEMISTRY　283　(1)　66-75　2008/01

DOI： 10.1074/jbc.M704686200 　

ISSN： 0021-9258
Conditional inactivation of Fbxw7 impairs cell-cycle exit during T cell differentiation and results in lymphomatogenesis. International-journal Peer-reviewed

Ichiro Onoyama, Ryosuke Tsunematsu, Akinobu Matsumoto, Taichi Kimura, Ignacio Moreno de Alborán, Keiko Nakayama, Keiichi I Nakayama

The Journal of experimental medicine　204　(12)　2875-88　2007/11/26

DOI： 10.1084/jem.20062299 　

eISSN： 1540-9538

More details Close

Cell proliferation is strictly controlled during differentiation. In T cell development, the cell cycle is normally arrested at the CD4(+)CD8(+) stage, but the mechanism underlying such differentiation-specific exit from the cell cycle has been unclear. Fbxw7 (also known as Fbw7, Sel-10, hCdc4, or hAgo), an F-box protein subunit of an SCF-type ubiquitin ligase complex, induces the degradation of positive regulators of the cell cycle, such as c-Myc, c-Jun, cyclin E, and Notch. FBXW7 is often mutated in a subset of human cancers. We have now achieved conditional inactivation of Fbxw7 in the T cell lineage of mice and found that the cell cycle is not arrested at the CD4(+)CD8(+) stage in the homozygous mutant animals. The mutant mice manifested thymic hyperplasia as a result of c-Myc accumulation and eventually developed thymic lymphoma. In contrast, mature T cells of the mutant mice failed to proliferate in response to mitogenic stimulation and underwent apoptosis in association with accumulation of c-Myc and p53. These latter abnormalities were corrected by deletion of p53. Our results suggest that Fbxw7 regulates the cell cycle in a differentiation-dependent manner, with its loss resulting in c-Myc accumulation that leads to hyperproliferation in immature T cells but to p53-dependent cell-cycle arrest and apoptosis in mature T cells.
NPKC epsilon, a P2Y(2)-R downstream effector in regulated mucin secretion from airway goblet cells Peer-reviewed

Camille Ehre, Yunxiang Zhu, Lubna H. Abdullah, John Olsen, Keiichi I. Nakayama, Keiko Nakayama, Robert O. Messing, C. William Davis

AMERICAN JOURNAL OF PHYSIOLOGY-CELL PHYSIOLOGY　293　(5)　C1445-C1454　2007/11

DOI： 10.1152/ajpcell.00051.2007 　

ISSN： 0363-6143
Ablation of p27 enhance kainate-induced seizure and hippocampal degeneration Peer-reviewed

Chihiro Ueyama, Hiroki Akashiba, Keiko Nakayama, Keiichi I. Nakayama, Nobuyoshi Nishiyama, Norio Matsuki

NEUROREPORT　18　(17)　1781-1785　2007/11

DOI： 10.1097/WNR.0b013e3282f16df6 　

ISSN： 0959-4965
Negative feedback loop in the Bim-caspase-3 axis regulating apoptosis and activity of osteoclasts Peer-reviewed

Hidetoshi Wakeyama, Toru Akiyama, Katsuhiko Takahashi, Hitoshi Amano, Yuho Kadono, Masaki Nakamura, Yasushi Oshima, Hiroyuki Itabe, Keiichi I. Nakayama, Keiko Nakayama, Kozo Nakamura, Sakae Tanaka

JOURNAL OF BONE AND MINERAL RESEARCH　22　(10)　1631-1639　2007/10

DOI： 10.1359/jbmr.070619 　

ISSN： 0884-0431
Essential role of Skp2-mediated p27 degradation in growth and adaptive expansion of pancreatic beta cells Peer-reviewed

Lingwen Zhong, Senta Georgia, Shuen-ing Tschen, Keiko Nakayama, Keiichi Nakayama, Anil Bhushan

JOURNAL OF CLINICAL INVESTIGATION　117　(10)　2869-2876　2007/10

DOI： 10.1172/JCI32198 　

ISSN： 0021-9738
Renal damage in obstructive nephropathy is decreased in Skp2-deficient mice Peer-reviewed

Sayuri Suzuki, Hirotaka Fukasawa, Kyoko Kitagawa, Chiharu Uchida, Takayuki Hattori, Tomoyasu Isobe, Toshiaki Oda, Taro Misaki, Naro Ohashi, Keiko Nakayama, Keiichi I. Nakayama, Akira Hishida, Tatsuo Yamamoto, Masatoshi Kitagawa

AMERICAN JOURNAL OF PATHOLOGY　171　(2)　473-483　2007/08

DOI： 10.2353/ajpath.2007.070279 　

ISSN： 0002-9440
Foxo3a is essential for maintenance of the hematopoietic stem cell pool Invited Peer-reviewed

Kana Miyamoto, Kiyomi Y. Araki, Kazuhito Naka, Fumio Arai, Keiyo Takubo, Satoshi Yamazaki, Sahoko Matsuoka, Takeshi Miyamoto, Keisuke Ito, Masako Ohmura, Chen Chen, Kentaro Hosokawa, Hiromitsu Nakauchi, Keiko Nakayama, Keiichi I. Nakayama, Mine Harada, Noboru Motoyama, Toshio Suda, Atsushi Hirao

CELL STEM CELL　1　(1)　101-112　2007/07

DOI： 10.1016/j.stem.2007.02.001 　

ISSN： 1934-5909
Cyclin D2 translocates p27 out of the nucleus and promotes its degradation at the G(0)-G(1) transition Peer-reviewed

Etsuo Susaki, Keiko Nakayama, Keiichi I. Nakayama

MOLECULAR AND CELLULAR BIOLOGY　27　(13)　4626-4640　2007/07

DOI： 10.1128/MCB.00862-06 　

ISSN： 0270-7306
The F box protein S phase kinase-associated protein 2 regulates adipose mass and adipocyte number in vivo Peer-reviewed

Paul S. Cooke, Denise R. Holsberger, Melissa A. Cimafranca, Daryl D. Meling, Charity M. Beals, Keiko Nakayama, Keiichi I. Nakayama, Hiroaki Kiyokawa

OBESITY　15　(6)　1400-1408　2007/06

DOI： 10.1038/oby.2007.168 　

ISSN： 1930-7381
Protein kinase C-delta phosphorylates Ebp1 and prevents its proteolytic degradation, enhancing cell survival Peer-reviewed

Zhixue Liu, Xia Liu, Keiichi I. Nakayama, Keiko Nakayama, Keqiang Ye

JOURNAL OF NEUROCHEMISTRY　100　(5)　1278-1288　2007/03

DOI： 10.1111/j.1471-4159.2006.04313.x 　

ISSN： 0022-3042
Asbestos-induced peribronchiolar cell proliferation and cytokine production are attenuated in lungs of protein kinase C-delta knockout mice

Arti Shukla, Karen M. Lounsbury, Trisha F. Barrett, Joanna Gell, Mercedes Rincon, Kelly J. Butnor, Douglas J. Taatjes, Gerald S. Davis, Pamela Vacek, Keiichi I. Nakayama, Keiko Nakayama, Chad Steele, Brooke T. Mossman

AMERICAN JOURNAL OF PATHOLOGY　170　(1)　140-151　2007/01

DOI： 10.2353/ajpath.2007.060381 　

ISSN： 0002-9440
Noncanonical Wnt signaling through G protein-linked PKC delta activation promotes bone formation

Xiaolin Tu, Kyu Sang Joeng, Keiichi I. Nakayama, Keiko Nakayama, Jayaraj Rajagopal, Thomas J. Carroll, Andrew P. McMahon, Fanxin Long

DEVELOPMENTAL CELL　12　(1)　113-127　2007/01

DOI： 10.1016/j.devcel.2006.11.003 　

ISSN： 1534-5807
The cyclin-dependent kinase inhibitors p57 and p27 regulate neuronal migration in the developing mouse neocortex

Yasuhiro Itoh, Norihisa Masuyama, Keiko Nakayama, Keiichi I. Nakayama, Yukiko Gotoh

JOURNAL OF BIOLOGICAL CHEMISTRY　282　(1)　390-396　2007/01

DOI： 10.1074/jbc.M609944200 　

ISSN： 0021-9258
Skp2 controls adipocyte proliferation during the development of obesity

Tamon Sakai, Hiroshi Sakaue, Takehiro Nakamura, Mitsuru Okada, Yasushi Matsuki, Eijiro Watanabe, Ryuji Hiramatsu, Keiko Nakayama, Keiichi I. Nakayama, Masato Kasuga

JOURNAL OF BIOLOGICAL CHEMISTRY　282　(3)　2038-2046　2007/01

DOI： 10.1074/jbc.M608144200 　

ISSN： 0021-9258
Up-regulation of GPR48 induced by down-regulation of p27(Kip1) enhances carcinoma cell invasiveness and metastasis

Yun Gao, Kyoko Kitagawa, Yoshihiro Hiramatsu, Hirotoshi Kikuchi, Tomoyasu Isobe, Mai Shimada, Chiharu Uchida, Takayuki Hattori, Toshiaki Oda, Keiko Nakayama, Keiichi I. Nakayama, Tatsuo Tanaka, Hiroyuki Konno, Masatoshi Kitagawa

CANCER RESEARCH　66　(24)　11623-11631　2006/12

DOI： 10.1158/0008-5472.CAN-06-2629 　

ISSN： 0008-5472
PKC delta regulates collagen-induced platelet aggregation through inhibition of VASP-mediated filopodia formation

Giordano Pula, Kai Schuh, Keiko Nakayama, Keiichi I. Nakayama, Ulrich Walter, Alastair W. Poole

BLOOD　108　(13)　4035-4044　2006/12

DOI： 10.1182/blood-2006-05-023739 　

ISSN： 0006-4971
Geminin is essential for the development of preimplantation mouse embryos

Kentaro Hara, Keiichi I. Nakayama, Keiko Nakayama

GENES TO CELLS　11　(11)　1281-1293　2006/11

DOI： 10.1111/j.1365-2443.2006.01019.x 　

ISSN： 1356-9597
Mitogenic signalling and the p16(INK4a)-Rb pathway cooperate to enforce irreversible cellular senescence

Akiko Takahashi, Naoko Ohtani, Kimi Yamakoshi, Shin-ichi Iida, Hidetoshi Tahara, Keiko Nakayama, Keiichi I. Nakayama, Toshinori Ide, Hideyuki Saya, Eiji Hara

NATURE CELL BIOLOGY　8　(11)　1291-U63　2006/11

DOI： 10.1038/ncb1491 　

ISSN： 1465-7392
[Ubiquitin system regulating G1 and S phases of cell cycle].

Keiichi I Nakayama, Keiko Nakayama

Tanpakushitsu kakusan koso. Protein, nucleic acid, enzyme　51　(10 Suppl)　1362-9　2006/08

ISSN： 0039-9450
HSP27 favors ubiquitination and proteasomal degradation of p27(Kip1) and helps S-phase re-entry in stressed cells Peer-reviewed

Arnaud Parcellier, Mathilde Brunet, Elise Schmitt, Edwige Col, Celine Didelot, Arlette Hammann, Keiko Nakayama, Keiichi I. Nakayama, Saadi Khochbin, Eric Solary, Carmen Garrido

FASEB JOURNAL　20　(8)　1179-+　2006/06

DOI： 10.1096/fj.05-4184fje 　

ISSN： 0892-6638
Double knockout mice show BASH and PKC delta have different epistatic relationships in B cell maturation and CD40-mediated activation Peer-reviewed

T Nojima, K Hayashi, R Goitsuka, K Nakayama, K Nakayama, D Kitamura

IMMUNOLOGY LETTERS　105　(1)　48-54　2006/05

DOI： 10.1016/j.imlet.2005.12.004 　

ISSN： 0165-2478
PKC delta is necessary for Smad3 expression and transforming growth factor beta-induced fibronectin synthesis in vascular smooth muscle cells Peer-reviewed

EJ Ryer, RP Hom, K Sakakibara, KI Nakayama, K Nakayama, PL Faries, B Liu, KC Kent

ARTERIOSCLEROSIS THROMBOSIS AND VASCULAR BIOLOGY　26　(4)　780-786　2006/04

DOI： 10.1161/01.ATV.0000209517.00220.cd 　

ISSN： 1079-5642

eISSN： 1524-4636
Suppression of centrosome amplification after DNA damage depends on p27 accumulation Peer-reviewed

E Sugihara, M Kanai, S Saito, T Nitta, H Toyoshima, K Nakayama, KI Nakayama, K Fukasawa, M Schwab, H Saya, M Miwa

CANCER RESEARCH　66　(8)　4020-4029　2006/04

DOI： 10.1158/0008-5472.CAN-05-3250 　

ISSN： 0008-5472
Two E3 ubiquitin ligases, SCF-Skp2 and DDB1-Cul4, target human Cdt1 for proteolysis Peer-reviewed

H Nishitani, N Sugimoto, Roukos, V, Y Nakanishi, M Saijo, C Obuse, T Tsurimoto, KI Nakayama, K Nakayama, M Fujita, Z Lygerou, T Nishimoto

EMBO JOURNAL　25　(5)　1126-1136　2006/03

DOI： 10.1038/sj,emboj.7601002 　

ISSN： 0261-4189
Identification of novel E2F1 target genes regulated in cell cycle-dependent and independent manners Peer-reviewed

R Iwanaga, H Komori, S Ishida, N Okamura, K Nakayama, KI Nakayama, K Ohtani

ONCOGENE　25　(12)　1786-1798　2006/03

DOI： 10.1038/sj.onc.1209210 　

ISSN： 0950-9232
Impaired germ cell development due to compromised cell cycle progression in Skp2-deficient mice Peer-reviewed

Abbas Fotovati, Keiko Nakayama, Keiichi I. Nakayama

CELL DIVISION　1　4-4　2006

DOI： 10.1186/1747-1028-1-4 　

ISSN： 1747-1028
Mammalian E4 is required for cardiac development and maintenance of the nervous system Peer-reviewed

C Kaneko-Oshikawa, T Nakagawa, M Yamada, H Yoshikawa, M Matsumoto, M Yada, S Hatakeyama, K Nakayama, KI Nakayama

MOLECULAR AND CELLULAR BIOLOGY　25　(24)　10953-10964　2005/12

DOI： 10.1128/MCB.25.24.10953-10964.2005 　

ISSN： 0270-7306
Phosphorylation of p27(KIP1) in the developing retina and retinoblastoma. Peer-reviewed

Kase Satoru, Yoshida Kazuhiko, Nakayama Keiichi I, Nakayama Keiko, Ikeda Hiromi, Harada Takayuki, Harada Chikako, Ohgami Kazuhiro, Shiratori Kenji, Ohno Shigeaki

Int J Mol Med　16　(2)　257-262　2005/08
Regulation of the cell cycle by SCF-type ubiquitin ligases. International-journal Peer-reviewed

Keiichi I Nakayama, Keiko Nakayama

Seminars in cell & developmental biology　16　(3)　323-33　2005/06

ISSN： 1084-9521

More details Close

Regulation of the cell cycle is dependent on protein degradation by the ubiquitin-proteasome system. Two major ubiquitin ligases, the anaphase-promoting complex or cyclosome (APC/C) and SCF complex, are responsible for the periodic proteolysis of many regulators of the cell cycle. The receptor component of the SCF complex is one of many F-box proteins, three of which--Skp2, Fbw7, and beta-TrCP--are well characterized and implicated in cell cycle regulation. We have generated mice deficient in Skp2, Fbw7, or beta-TrCP1 and have identified the roles of these proteins in both cell cycle regulation and mouse development. Clinical evidence also suggests that dysregulation of these F-box proteins contributes to human cancers.
Disappearance of p27(KIP1) and increase in proliferation of the lens cells after extraction of most of the fiber cells of the lens Peer-reviewed

S Kase, K Yoshida, H Ikeda, T Harada, C Harada, J Imaki, K Ohgami, K Shiratori, KI Nakayama, K Nakayama, S Ohno

CURRENT EYE RESEARCH　30　(6)　437-442　2005/06

DOI： 10.1080/02713680590959286 　

ISSN： 0271-3683
Ubiquitylation of RAG-2 by Skp2-SCF links destruction of the V(D)J recombinase to the cell cycle Peer-reviewed

H Jiang, FC Chang, AE Ross, JY Lee, K Nakayama, K Nakayama, S Desiderio

MOLECULAR CELL　18　(6)　699-709　2005/06

DOI： 10.1016/j.molcel.2005.05.011 　

ISSN： 1097-2765
Bcl-2-related protein A1 is an endogenous and cytokine-stimulated mediator of cytoprotection in hyperoxic acute lung injury. Peer-reviewed

He Chuan Hua, Waxman Aaron B, Lee Chun Geun, Link Holger, Rabach Morgan E, Ma Bing, Chen Qingsheng, Zhu Zhou, Zhong Mei, Nakayama Keiko, Nakayama Keiichi I, Homer Robert, Elias Jack A

J Clin Invest　115　(4)　1039-1048　2005/04

DOI： 10.1172/JCI200523004 　
Suppression of cell migration by protein kinase C delta Peer-reviewed

D Jackson, Y Zheng, DG Lyo, YJ Shen, K Nakayama, KI Nakayama, MJ Humphries, ME Reyland, DA Foster

ONCOGENE　24　(18)　3067-3072　2005/04

DOI： 10.1038/sj.onc.1208465 　

ISSN： 0950-9232
p107 inhibits G1 to S phase progression by downregulating expression of the F-box protein Skp2 Peer-reviewed

G Rodier, C Makris, P Coulombe', A Scime, K Nakayama, KI Nakayama, S Meloche

JOURNAL OF CELL BIOLOGY　168　(1)　55-66　2005/01

DOI： 10.1083/jcb.200404146 　

ISSN： 0021-9525
Role of serine 10 phosphorylation in p27 stabilization revealed by analysis of p27 knock-in mice harboring a serine 10 mutation Peer-reviewed

Y Kotake, K Nakayama, N Ishida, KI Nakayama

JOURNAL OF BIOLOGICAL CHEMISTRY　280　(2)　1095-1102　2005/01

DOI： 10.1074/jbc.M406117200 　

ISSN： 0021-9258
Functional regulation of FEZ1 by the U-box-type ubiquitin ligase E4B contributes to neuritogenesis. International-journal Peer-reviewed

Okumura, F, Hatakeyama, S, Matsumoto, M, Kamura, T, Nakayama, K.I

Journal of Biological Chemistry　279　(51)　53533-53543-43　2004/12

ISSN： 0021-9258

More details Close

E4B (also known as UFD2a) is a mammalian homolog of Saccharomyces cerevisiae Ufd2, which was originally described as a ubiquitin chain assembly factor (E4). E4B is a U-box-type ubiquitin-protein isopeptide ligase (E3) and likely functions as either an E3 or an E4. With a yeast two-hybrid screen, we have now identified FEZ1 (fasciculation and elongation protein zeta 1) as a protein that interacts with E4B. FEZ1 is implicated in neuritogenesis when phosphorylated by protein kinase Czeta (PKCzeta). Interaction between E4B and FEZ1 in mammalian cells was enhanced by coexpression of constitutively active PKCzeta. E4B mediated the polyubiquitylation of FEZ1 but did not affect its intracellular stability, suggesting that such modification of FEZ1 is not a signal for its proteolysis. Polyubiquitylation of FEZ1 by E4B required Lys(27) of ubiquitin. Expression of a dominant-negative mutant of E4B in rat pheochromocytoma PC12 cells resulted in inhibition of neurite extension induced either by nerve growth factor or by coexpression of FEZ1 and constitutively active PKCzeta. These findings indicate that E4B serves as a ubiquitin ligase for FEZ1 and thereby regulates its function but not its degradation.
Cytoplasmic ubiquitin ligase KPC regulates proteolysis of p27(Kip1) at G1 phase Peer-reviewed

T Kamura, T Hara, M Matsumoto, N Ishida, F Okumura, S Hatakeyama, M Yoshida, K Nakayama, KI Nakayama

NATURE CELL BIOLOGY　6　(12)　1229-1235　2004/12

DOI： 10.1038/ncb1194 　

ISSN： 1465-7392
The ISG15 isopeptidase UBP43 is regulated by proteolysis via the SCFSkp2 ubiquitin ligase Peer-reviewed

S Tokarz, C Berset, J La Rue, K Friedman, KI Nakayama, K Nakayama, DE Zhang, S Lanker

JOURNAL OF BIOLOGICAL CHEMISTRY　279　(45)　46424-46430　2004/11

DOI： 10.1074/jbc.M403189200 　

ISSN： 0021-9258
Early embryonic death in mice lacking the beta-catenin-binding protein duplin Peer-reviewed

M Nishiyama, K Nakayama, R Tsunematsu, T Tsukiyama, A Kikuchi, KI Nakayama

MOLECULAR AND CELLULAR BIOLOGY　24　(19)　8386-8394　2004/10

DOI： 10.1128/MCB.24.19.8386-8394.2004 　

ISSN： 0270-7306
【癌から発生・分化,さらに再生医学へ拡大する細胞周期研究】細胞周期制御分子の発生過程に果たす役割遺伝子操作マウスは何を語るか?

中山啓子, 中山敬一

実験医学　22　(13)　1814-1819　2004/09
Abnormal angiogenesis in Foxo1 (Fkhr)-deficient mice. International-journal

Tatsuo Furuyama, Kazuko Kitayama, Yuri Shimoda, Minetaro Ogawa, Kiyoaki Sone, Kiyomi Yoshida-Araki, Hiroshi Hisatsune, Shin-ichi Nishikawa, Keiko Nakayama, Keiichi Nakayama, Kyoji Ikeda, Noboru Motoyama, Nozomu Mori

The Journal of biological chemistry　279　(33)　34741-9　2004/08/13

ISSN： 0021-9258

More details Close

Members of the Foxo family, Foxo1 (Fkhr), Foxo3 (Fkhrl1), and Foxo4 (Afx), are mammalian homologs of daf-16, which influences life span and energy metabolism in Caenorhabditis elegans. Mammalian FOXO proteins also play important roles in cell cycle arrest, apoptosis, stress resistance, and energy metabolism. In this study, we generated Foxo1-deficient mice to investigate the physiological role of FOXO1. The Foxo1-deficient mice died around embryonic day 11 because of defects in the branchial arches and remarkably impaired vascular development of embryos and yolk sacs. In vitro differentiation of embryonic stem cells demonstrated that endothelial cells derived from wild-type and Foxo1-deficient embryonic stem cells were able to produce comparable numbers of colonies supported by a layer of OP9 stromal cells. Although the morphology of the endothelial cell colonies was identical in both genotypes in the absence of exogenous vascular endothelial growth factor (VEGF), Foxo1-deficient endothelial cells showed a markedly different morphological response compared with wild-type endothelial cells in the presence of exogenous VEGF. These results suggest that Foxo1 is essential to the ability of endothelial cells to respond properly to a high dose of VEGF, thereby playing a critical role in normal vascular development.
Distinct roles of c-Abl and Atm in oxidative stress response are mediated by protein kinase C delta Peer-reviewed

BJ Li, XY Wang, N Rasheed, YY Hu, S Boast, T Ishii, K Nakayama, KI Nakayama, SP Goff

GENES & DEVELOPMENT　18　(15)　1824-1837　2004/08

DOI： 10.1101/gad.1223504 　

ISSN： 0890-9369
Involvement of p27(KIP1) in the proliferation of the developing corneal endothelium Peer-reviewed

K Yoshida, S Kase, K Nakayama, H Nagahama, T Harada, H Ikeda, C Harada, J Imaki, K Ohgami, K Shiratori, IB Ilieva, S Ohno, S Nishi, KI Nakayama

INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE　45　(7)　2163-2167　2004/07

DOI： 10.1167/iovs.03-1238 　

ISSN： 0146-0404
Phosphorylation-dependent degradation of c-Myc is mediated by the F-box protein Fbw7. International-journal Peer-reviewed

Masayoshi Yada, Shigetsugu Hatakeyama, Takumi Kamura, Masaaki Nishiyama, Ryosuke Tsunematsu, Hiroyuki Imaki, Noriko Ishida, Fumihiko Okumura, Keiko Nakayama, Keiichi I Nakayama

The EMBO journal　23　(10)　2116-25　2004/05/19

DOI： 10.1038/sj.emboj.7600217 　

ISSN： 0261-4189

More details Close

The F-box protein Skp2 mediates c-Myc ubiquitylation by binding to the MB2 domain. However, the turnover of c-Myc is largely dependent on phosphorylation of threonine-58 and serine-62 in MB1, residues that are often mutated in cancer. We now show that the F-box protein Fbw7 interacts with and thereby destabilizes c-Myc in a manner dependent on phosphorylation of MB1. Whereas wild-type Fbw7 promoted c-Myc turnover in cells, an Fbw7 mutant lacking the F-box domain delayed it. Furthermore, depletion of Fbw7 by RNA interference increased both the abundance and transactivation activity of c-Myc. Accumulation of c-Myc was also apparent in mouse Fbw7-/- embryonic stem cells. These observations suggest that two F-box proteins, Fbw7 and Skp2, differentially regulate c-Myc stability by targeting MB1 and MB2, respectively.
Skp2-mediated degradation of p27 regulates progression into mitosis Peer-reviewed

K Nakayama, H Nagahama, YA Minamishima, S Miyake, N Ishida, S Hatakeyama, M Kitagawa, S Iemura, T Natsume, KI Nakayama

DEVELOPMENTAL CELL　6　(5)　661-672　2004/05

DOI： 10.1016/S1534-5807(04)00131-5 　

ISSN： 1534-5807

eISSN： 1878-1551
The papillomavirus E7 oncoprotein is ubiquitinated by UbcH7 and Cullin 1- and Skp2-containing E3 ligase Peer-reviewed

KJ Oh, A Kalinina, J Wang, K Nakayama, KI Nakayama, S Bagchi

JOURNAL OF VIROLOGY　78　(10)　5338-5346　2004/05

DOI： 10.1128/JVI.78.10.5338-5346.2004 　

ISSN： 0022-538X
Distribution of p27(KIP1), cyclin D1, and proliferating cell nuclear antigen after retinal detachment Peer-reviewed

K Yoshida, S Kase, K Nakayama, H Nagahama, T Harada, H Ikeda, C Harada, J Imaki, K Ohgami, K Shiratori, S Ohno, KI Nakayama

GRAEFES ARCHIVE FOR CLINICAL AND EXPERIMENTAL OPHTHALMOLOGY　242　(5)　437-441　2004/05

DOI： 10.1007/s00417-004-0861-7 　

ISSN： 0721-832X
Involvement of p27(KIP1) in proliferation of the retinal pigment epithelium and ciliary body Peer-reviewed

K Yoshida, K Nakayama, S Kase, H Nagahama, T Harada, H Ikeda, C Harada, J Imaki, K Ohgami, K Shiratori, S Ohno, S Nishi, KI Nakayama

ANATOMY AND EMBRYOLOGY　208　(2)　145-150　2004/05

DOI： 10.1007/s00429-004-0382-5 　

ISSN： 0340-2061
Mouse Fbw7/Sel-10/Cdc4 is required for notch degradation during vascular development. International-journal Peer-reviewed

Ryosuke Tsunematsu, Keiko Nakayama, Yuichi Oike, Masaaki Nishiyama, Noriko Ishida, Shigetsugu Hatakeyama, Yasumasa Bessho, Ryoichiro Kageyama, Toshio Suda, Keiichi I Nakayama

The Journal of biological chemistry　279　(10)　9417-23　2004/03/05

DOI： 10.1074/jbc.M312337200 　

ISSN： 0021-9258

More details Close

Mammalian Fbw7 (also known as Sel-10, hCdc4, or hAgo) is the F-box protein component of an SCF (Skp1-Cul1-F-box protein-Rbx1)-type ubiquitin ligase, and the mouse Fbw7 is expressed prominently in the endothelial cell lineage of embryos. We generated mice deficient in Fbw7 and found that the embryos died in utero at embryonic day 10.5-11.5, manifesting marked abnormalities in vascular development. Vascular remodeling was impaired in the brain and yolk sac, and the major trunk veins were not formed. In vitro para-aortic splanchnopleural explant cultures from Fbw7(-/-) embryos also manifested an impairment of vascular network formation. Notch4, which is the product of the proto-oncogene Int3 and an endothelial cell-specific mammalian isoform of Notch, accumulated in Fbw7(-/-) embryos, resulting in an increased expression of Hey1, which encodes a transcriptional repressor that acts downstream of Notch signaling and is implicated in vascular development. Expression of Notch1, -2, or -3 or of cyclin E was unaffected in Fbw7(-/-) embryos. Mammalian Fbw7 thus appears to play an indispensable role in negative regulation of the Notch4-Hey1 pathway and is required for vascular development.
マウス網膜色素上皮及び毛様体の細胞増殖制御におけるp27(KIP1)の役割

加瀬諭, 吉田和彦, 大野重昭, 中山啓子, 中山敬一

日本眼科学会雑誌　108　(臨増)　205-205　2004/03
Association of cathepsin E deficiency with development of atopic dermatitis. International-journal Peer-reviewed

Tsukuba Takayuki, Okamoto Kuniaki, Okamoto Yoshiko, Yanagawa Michiyo, Kohmura Keiko, Yasuda Yoshiyuki, Uchi Hiroshi, Nakahara Takeshi, Furue Masutaka, Nakayama Keiko, Kadowaki Tomoko, Yamamoto Kenji, Nakayama Keiichi I

J Biochem (Tokyo)　134　(6)　893-902　2003/12

DOI： 10.1093/jb/mvg216 　

ISSN： 0021-924X

More details Close

Atopic dermatitis (AD) is a pruritic inflammatory skin diseases associated with a family history of atropy. Here we show that mice lacking the endolysosomal aspartic proteinase cathepsin E spontaneously develop skin lesions similar to those of humans with AD when reared under conventional conditions but not under specific pathogen-free conditions. These mice showed the increase in the ratio of CD4+/CD8+ T cells, the strong polarization of naïve T cells to T helper 2 cells, and the systemic accumulation of IL-18 and IL-1beta accompanied by a marked increase in IL-4, IL-5, and IgE. The relative rates of degradation of IL-18 and IL-1beta were significantly lower in cathepsin E-deficient mice than wild-type mice. These results strongly suggest that the development of AD in cathepsin E-deficient mice is initiated by systemic accumulation of IL-18 and IL-1beta, mainly due to their reduced turnover rates. In addition, the reduced expression of cathepsin E was also observed in erythrocytes of both humans with AD and the AD mouse model NC/Nga. Cathepsin E deficiency might thus be responsible for the induction of AD in humans and mice.
CDKインヒビターp27の機能と制御

中山啓子

Proceedings of Clinical Electron Microscopy　25　7-11　2003/12
医学・医療の進歩を世界へ向けて発生分化・生殖医療発生分化発生分化研究におけるモデル動物

中山啓子

日本医学会総会26回会誌　(2)　164-164　2003/12
PKC-δノックアウトマウスにおけるB細胞の過剰増殖と自己免疫疾患

中山啓子

東北医学雑誌　115　(2)　145-147　2003/12
Publisher:
ISSN： 0040-8700
角膜上皮剥離後及び発生過程の水晶体の細胞増殖における細胞周期調節蛋白の役割

吉田和彦, 原田高幸, 原田知加子, 加瀬諭, 池田裕美, 酒井正春, 西信三, 今城純子, 中山啓子, 永濱裕康, 中山敬一, 大野重昭

日本眼科学会雑誌　107　(11)　678-686　2003/11

ISSN： 0029-0203
P57(KIP2) modulates stress-activated signaling by inhibiting c-Jun NH2-terminal kinase/stress-activated protein kinase Peer-reviewed

TS Chang, MJ Kim, K Ryoo, J Park, SJ Eom, J Shim, KI Nakayama, K Nakayama, M Tomita, K Takahashi, MJ Lee, EJ Choi

JOURNAL OF BIOLOGICAL CHEMISTRY　278　(48)　48092-48098　2003/11

DOI： 10.1074/jbc.M309421200 　

ISSN： 0021-9258
Degradation of p57(Kip2) mediated by SCFSkp2 - dependent ubiquitylation Peer-reviewed

T Kamura, T Hara, S Kotoshiba, M Yada, N Ishida, H Imaki, S Hatakeyama, K Nakayama, KI Nakayama

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA　100　(18)　10231-10236　2003/09

DOI： 10.1073/pnas.1831009100 　

ISSN： 0027-8424
HDACに結合する分化抑制因子EIDファミリーの機能解析

三宅智, 笹島由加, 柳澤夕佳, 中山啓子, 湯浅保仁

日本癌学会62回総会記事　256-257　2003/08
ノックインマウスを用いたp27セリン10残基リン酸化についての検討

中山啓子, 中山敬一

日本癌学会62回総会記事　137-137　2003/08
Cell cycle-dependent regulation of the Skp2 promoter by GA-binding protein Peer-reviewed

H Imaki, K Nakayama, S Delehouzee, H Handa, M Kitagawa, T Kamura, KI Nakayama

CANCER RESEARCH　63　(15)　4607-4613　2003/08

ISSN： 0008-5472
Role of the SCFSkp2 ubiquitin ligase in the degradation of p21(Cip1) in S phase Peer-reviewed

G Bornstein, J Bloom, D Sitry-Shevah, K Nakayama, M Pagano, A Hershko

JOURNAL OF BIOLOGICAL CHEMISTRY　278　(28)　25752-25757　2003/07

DOI： 10.1074/jbc.M301774200 　

ISSN： 0021-9258
Impaired degradation of inhibitory subunit of NF-kappa B (I kappa B) and beta-catenin as a result of targeted disruption of the beta-TrCP1 gene Peer-reviewed

K Nakayama, S Hatakeyama, S Maruyama, A Kikuchi, K Onoe, RA Good, KI Nakayama

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA　100　(15)　8752-8757　2003/07

DOI： 10.1073/pnas.1133216100 　

ISSN： 0027-8424
The F-box protein Skp2 participates in c-Myc proteosomal degradation and acts as a cofactor for c-Myc-regulated transcription. Peer-reviewed

von der Lehr Natalie, Johansson Sara, Wu Siqin, Bahram Fuad, Castell Alina, Cetinkaya Cihan, Hydbring Per, Weidung Ingrid, Nakayama Keiko, Nakayama Keiichi I, Soderberg Ola, Kerppola Tom K, Larsson Lars-Gunnar

Mol Cell　11　(5)　1189-1200　2003/05

DOI： 10.1016/S1097-2765(03)00193-X 　
The role of p57Kip2 on mandibular growth in mice: By means of laser microdissection for hard tissues Peer-reviewed

Watahiki J, Yamaguchi T, Irie T, Takahashi K, Nakano H, Yagasaki Y, Nakayama K. I, Nakayama K, Maki K, Tachikawa T

Orthod. Waves　62　(3)　201-206　2003/04
Publisher:
ISSN： 1344-0241

More details Close

p57^<KiP2> is a member of the cip/kip family of negative regulators of the cell cycle and is strongly expressed in developing chondrocytes. In ^<KiP2>-deficient mice, shortening of limbs caused by delay of calcification in endochondral ossification has been confirmed as a skeletal abnormality. The role of p57^<KiP2> has been unclear in maxillofacial morphology. In this study to evaluate the involvement of p57^<KiP2> in mandibular growth during postnatal growth, we examined the expression of p57^<KiP2> in the mandibular condylar cartilage. Expression was confirmed in the proliferative and mature-cell layers; the strongest expression was in the mature-cell layer. We also examined the expression of type X collagen genes in p57^<KiP2>-deficient mice by real time PCR and found that the expression was much lower in the condylar cartilage of newborn p57^<KiP2>-deficient mice than in that of newborn wild-type mice. It is, therefore, concluded that ^<KiP2> may play an important role in mandibular growth by promoting the final differentiation of chondrocytes in the mandibular condylar cartilage.
A novel route for connexin 43 to inhibit cell proliferation: Negative regulation of S-Phase kinase-associated protein (Skp 2) Peer-reviewed

YW Zhang, K Nakayama, K Nakayama, Morita, I

CANCER RESEARCH　63　(7)　1623-1630　2003/04

ISSN： 0008-5472
Characterization of the mouse gene for the U-box-type ubiquitin lipase UFD2a Peer-reviewed

C Kaneko, S Hatakeyama, M Matsumoto, M Yada, K Nakayama, KI Nakayama

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS　300　(2)　297-304　2003/01

DOI： 10.1016/S0006-291X(02)02834-6 　

ISSN： 0006-291X
Glomerular differentiation in p27 and p57 double-mutant metanephroi Peer-reviewed

S Tomari, H Nagahama, YJ Shu, S Hoshi, K Nakayama, KI Nakayama, M Nagata

ANATOMY AND EMBRYOLOGY　206　(1-2)　31-36　2002/12

DOI： 10.1007/s00429-002-0274-5 　

ISSN： 0340-2061
Diminished alcohol preference in transgenic mice lacking aldehyde dehydrogenase activity

Toyohi Isse, Tsunehiro Oyama, Kyoko Kitagawa, Koji Matsuno, Akiko Matsumoto, Akira Yoshida, Keiko Nakayama, Kei-Ichi Nakayama, Toshihiro Kawamoto

Pharmacogenetics　12　(8)　621-626　2002/11

DOI： 10.1097/00008571-200211000-00006 　

ISSN： 0960-314X
PKC-δ遺伝子欠損マウスにおけるB細胞の過剰増殖と自己寛容の破綻

宮本顕友, 中山啓子, 広瀬幸子, 大野茂夫, 畠山鎮次, 中山敬一

日本免疫学会総会・学術集会記録　32　92-92　2002/10
Aplidin (TM) induces the mitochondrial apoptotic pathway via oxidative stress-mediated JNK and p38 activation and protein kinase C delta Peer-reviewed

LF Garcia-Fernandez, A Losada, Alcaide, V, AM Alvarez, A Cuadrado, L Gonzalez, K Nakayama, KI Nakayama, JM Fernandez-Sousa, A Munoz, JM Sanchez-Puelles

ONCOGENE　21　(49)　7533-7544　2002/10

DOI： 10.1038/sj.onc.1205972 　

ISSN： 0950-9232
Clinical and biological significance of S-phase kinase-associated protein 2 (Skp2) gene expression in gastric carcinoma: modulation of malignant phenotype by Skp2 overexpression, possibly via p27 proteolysis Peer-reviewed

T Masuda, H Inoue, H Sonoda, S Mine, Y Yoshikawa, K Nakayama, KI Nakayama, M Mori

CANCER RESEARCH　62　(13)　3819-3825　2002/07

ISSN： 0008-5472
Phosphorylation of p27Kip1 on serine 10 is required for its binding to CRM1 and nuclear export. International-journal Peer-reviewed

Noriko Ishida, Taichi Hara, Takumi Kamura, Minoru Yoshida, Keiko Nakayama, Keiichi I Nakayama

The Journal of biological chemistry　277　(17)　14355-8　2002/04/26

DOI： 10.1074/jbc.C100762200 　

ISSN： 0021-9258
TGF β receptor II型及び/或はp27マウスの標的撹乱におけるestrogen誘発下垂体腫瘍形成の形態的分子的分析

池田秀敏, 吉本高志, 志田直樹, 三好一郎, 中山啓子, 中山敬一, 大島正伸, 武藤誠

日本内分泌学会雑誌　78　(1)　103-103　2002/04
Skp2を介したp27(KIP1)の分解による角膜上皮細胞の増殖制御

吉田和彦, 山本和幸, 松田彰, 大野重昭, 中山啓子, 永濱裕康, 中山敬一

日本眼科学会雑誌　106　(臨増)　99-99　2002/04
Increased proliferation of B cells and auto-immunity in mice lacking protein kinase C delta Peer-reviewed

A Miyamoto, K Nakayama, H Imaki, S Hirose, Y Jiang, M Abe, T Tsukiyama, H Nagahama, S Ohno, S Hatakeyama, KI Nakayama

NATURE　416　(6883)　865-869　2002/04

DOI： 10.1038/416865a 　

ISSN： 0028-0836
アセトアルデヒド脱水素酵素(Aldh)2ノックアウトマウスは低アルコール嗜好性を示す

一瀬豊日, 北川恭子, 松本明子, 吉田昭, 川本俊弘, 中山啓子, 中山啓一

産業医科大学雑誌　24　(1)　94-94　2002/03
Publisher:
ISSN： 0387-821X
Role of the PLC-related, catalytically inactive protein p130 in GABA(A) receptor function Peer-reviewed

T Kanematsu, IS Jang, T Yamaguchi, H Nagahama, K Yoshimura, K Hidaka, M Matsuda, H Takeuchi, Y Misumi, K Nakayama, T Yamamoto, N Akaike, M Hirata, K Nakayama

EMBO JOURNAL　21　(5)　1004-1011　2002/03

DOI： 10.1093/emboj/21.5.1004 　

ISSN： 0261-4189
Involvement of p27(KIP1) degradation by Skp2 in the regulation of proliferation in response to wounding of corneal epithelium Peer-reviewed

K Yoshida, K Nakayama, H Nagahama, T Harada, C Harada, J Imaki, A Matsuda, K Yamamoto, M Ito, S Ohno, K Nakayama

INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE　43　(2)　364-370　2002/02

ISSN： 0146-0404
Recovery of liver mass without proliferation of hepatocytes after partial hepatectomy in skp2-deficient mice Peer-reviewed

YA Minamishima, K Nakayama, KI Nakayama

CANCER RESEARCH　62　(4)　995-999　2002/02

ISSN： 0008-5472
Interaction between cyclin T1 and SCFSKP2 targets CDK9 for ubiquitination and degradation by the proteasome Peer-reviewed

RE Kiernan, S Emiliani, E Nakayama, A Castro, JC Labbe, T Lorca, K Nakayama, A Benkirane

MOLECULAR AND CELLULAR BIOLOGY　21　(23)　7956-7970　2001/12

DOI： 10.1128/MCB.21.23.7956-7970.2001 　

ISSN： 0270-7306
Characterization of a mouse gene (Fbxw6) that encodes a homologue of Caenorhabditis elegans SEL-10 Peer-reviewed

S Maruyama, S Hatakeyama, K Nakayama, N Ishida, K Kawakami, K Nakayama

GENOMICS　78　(3)　214-222　2001/12

DOI： 10.1006/geno.2001.6658 　

ISSN： 0888-7543
Degradation of p27(Kip1) at the G(0)-G(1) transition mediated by a Skp2-independent ubiquitination pathway Peer-reviewed

T Hara, T Kamura, K Nakayama, K Oshikawa, S Hatakeyama, KI Nakayama

JOURNAL OF BIOLOGICAL CHEMISTRY　276　(52)　48937-48943　2001/12

DOI： 10.1074/jbc.M107274200 　

ISSN： 0021-9258
Morphologic and molecular analysis of estrogen-induced pituitary tumorigenesis in targeted disruption of transforming growth factor-beta receptor type II and/or p27 mice Peer-reviewed

H Ikeda, T Yoshimoto, N Shida, Miyoshi, I, K Nakayama, K Nakayama, M Oshima, MM Taketo

ENDOCRINE　16　(1)　55-65　2001/10

DOI： 10.1385/ENDO:16:1:55 　

ISSN： 0969-711X
胃癌におけるユビキチンリガーゼSCFSkp2の発現とその意義

増田隆明, 山下継史, 園田英人, 峯真司, 中山啓子, 中山敬一, 井上裕, 森正樹

日本癌学会60回総会記事　60回　96-96　2001/09
Publisher:
ISSN： 0546-0476
caspase-3ノックアウトマウスにおける難聴

中川尚志, 牧嶋知子, 金子千恵, 小宮山荘太郎, 中山敬一, 中山啓子

Audiology Japan　44　(5)　357-358　2001/09
Publisher:
DOI： 10.4295/audiology.44.357 　

ISSN： 0303-8106
遺伝子組換え動物モデルの新展開ユビキチンリガーゼ群ノックアウトマウスによる生体機能の解析特に細胞周期研究を中心として

中山敬一, 嘉村巧, 原太一, 畠山鎮次, 中山啓子

日本癌学会60回総会記事　30-30　2001/09
A mouse knock-in model exposes sequential proteolytic pathways that regulate p27(Kip1) in G1 and S phase Peer-reviewed

NP Malek, H Sundberg, S McGrew, K Nakayama, TR Kyriakidis, JM Roberts

NATURE　413　(6853)　323-327　2001/09

DOI： 10.1038/35095083 　

ISSN： 0028-0836
Deafness due to degeneration of cochlear neurons in caspase-3-deficient mice Peer-reviewed

H Morishita, T Makishima, C Kaneko, YS Lee, N Segil, K Takahashi, A Kuraoka, T Nakagawa, J Nabekura, K Nakayama, KI Nakayama

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS　284　(1)　142-149　2001/06

DOI： 10.1006/bbrc.2001.4939 　

ISSN： 0006-291X
Skp2を介した角膜上皮細胞の増殖制御

吉田和彦, 山本和之, 松田彰, 大野重昭, 中山啓子, 永濱裕康, 中山敬一

日本眼科学会雑誌　105　(臨増)　188-188　2001/03
Spatial and temporal expression patterns of the cyclin-dependent kinase (CDK) inhibitors p27(Kip1) and p57(Kip2) during mouse development Peer-reviewed

H Nagahama, S Hatakeyama, K Nakayama, M Nagata, K Tomita, K Nakayama

ANATOMY AND EMBRYOLOGY　203　(2)　77-87　2001/02

ISSN： 0340-2061
Down-regulation of p27(Kip1) expression is required for development and function of T cells Peer-reviewed

T Tsukiyama, N Ishida, M Shirane, YA Minamishima, S Hatakeyama, M Kitagawa, K Nakayama, K Nakayama

JOURNAL OF IMMUNOLOGY　166　(1)　304-312　2001/01

DOI： 10.4049/jimmunol.166.1.304 　

ISSN： 0022-1767
Hyperinsulinemia in PRIP-1 gene deleted mice Peer-reviewed

Naoko Doira, Takashi Kanematsu, Miho Matsuda, Hiroshi Takeuchi, Hito-O. Nakano, Yushi Ito, Keiko Nakayama, Kei-Ichi Nakayama, Masato Hirata

Biomedical Research　22　(3)　157-165　2001
Publisher: Biomedical Research Foundation
DOI： 10.2220/biomedres.22.157 　

ISSN： 1880-313X 0388-6107
NF-κBシグナル及び細胞周期制御に関与するユビキチンリガーゼ FWD1とSkp2

畠山鎮次, 中山啓子, 中山敬一

日本免疫学会総会・学術集会記録　30　221-221　2000/11
Chk1欠損マウスを用いたマウスChk1キナーゼの機能解析

高井裕之, 冨永薫, 本山昇, 池田恭治, 中西真, 永濱裕康, 中山啓子, 中山敬一

日本臨床分子医学会記録　37　70-70　2000/11
Tyk2 plays a restricted role in IFN alpha signaling, although it is required for IL-12-mediated T cell function Peer-reviewed

K Shimoda, K Kato, K Aoki, T Matsuda, A Miyamoto, M Shibamori, M Yamashita, A Numata, K Takase, S Kobayashi, S Shibata, Y Asano, H Gondo, K Sekiguchi, K Nakayama, T Nakayama, T Okamura, S Okamura, Y Niho, K Nakayama

IMMUNITY　13　(4)　561-571　2000/10

ISSN： 1074-7613
Rb経路の上流に位置するサイクリンEとp27Kip1の蛋白分解機構

中山敬一, 畠山鎮次, 北川雅敏, 小南欽一郎, 中山啓子

日本癌学会59回総会記事　419-419　2000/09
染色体倍数性及び中心体複製に異常をきたすノックアウトマウスの解析

中山啓子, 畠山鎮次, 北川雅敏, 小南欽一郎, 中山敬一

日本癌学会59回総会記事　422-422　2000/09
Phosphorylation at serine 10, a major phosphorylation site of p27(Kip1), increases its protein stability Peer-reviewed

N Ishida, M Kitagawa, S Hatakeyama, K Nakayama

JOURNAL OF BIOLOGICAL CHEMISTRY　275　(33)　25146-25154　2000/08

DOI： 10.1074/jbc.M001144200 　

ISSN： 0021-9258
Aldehyde dehydrogenase (ALDH) 2 associates with oxidation of methoxyacetaldehyde; in vitro analysis with liver subcellular fraction derived from human and Aldh2 gene targeting mouse Peer-reviewed

K Kitagawa, T Kawamoto, N Kunugita, T Tsukiyama, K Okamoto, A Yoshida, K Nakayama, K Nakayama

FEBS LETTERS　476　(3)　306-311　2000/07

DOI： 10.1016/S0014-5793(00)01710-5 　

ISSN： 0014-5793
Aberrant cell cycle checkpoint function and early embryonic death in Chk1(-/-) mice Peer-reviewed

H Takai, K Tominaga, N Motoyama, YA Minamishima, H Nagahama, T Tsukiyama, K Ikeda, K Nakayama, N Nakanishi, K Nakayama

GENES & DEVELOPMENT　14　(12)　1439-1447　2000/06

ISSN： 0890-9369

eISSN： 1549-5477
Targeted disruption of Skp2 results in accumulation of cyclin E and p27(Kip1), polyploidy and centrosome overduplication Peer-reviewed

K Nakayama, H Nagahama, YA Minamishima, M Matsumoto, Nakamichi, I, K Kitagawa, M Shirane, R Tsunematsu, T Tsukiyama, N Ishida, M Kitagawa, K Nakayama, S Hatakeyama

EMBO JOURNAL　19　(9)　2069-2081　2000/05

DOI： 10.1093/emboj/19.9.2069 　

ISSN： 0261-4189

eISSN： 1460-2075
Mice lacking a CDK inhibitor, p57(Kip2), exhibit skeletal abnormalities and growth retardation Peer-reviewed

K Takahashi, K Nakayama, K Nakayama

JOURNAL OF BIOCHEMISTRY　127　(1)　73-83　2000/01

ISSN： 0021-924X
IκBαとβ-cateninの分解に関与するユビキチン化酵素複合体SCFFWD1

畠山鎮次, 中山啓子, 中野裕康, 奥村康, 菊池章, 小野江和則, 中山敬一

日本免疫学会総会・学術集会記録　29　104-104　1999/10
Common pathway for the ubiquitination of I kappa B alpha, I kappa B beta, and I kappa B epsilon mediated by the F-box protein FWD1 Peer-reviewed

M Shirane, S Hatakeyama, K Hattori, K Nakayama, K Nakayama

JOURNAL OF BIOLOGICAL CHEMISTRY　274　(40)　28169-28174　1999/10

DOI： 10.1074/jbc.274.40.28169 　

ISSN： 0021-9258
β-カテニンのユビキチン化に関わる分子FWD1の同定と機能解析

北川雅敏, 畠山鎮次, 菊池章, 中山啓子, 中山敬一

日本癌学会58回総会記事　79-79　1999/08
An F-box protein, FWD1, mediates ubiquitin-dependent proteolysis of beta-catenin Peer-reviewed

M Kitagawa, S Hatakeyama, M Shirane, M Matsumoto, N Ishida, K Hattori, Nakamichi, I, A Kikuchi, K Nakayama, K Nakayama

EMBO JOURNAL　18　(9)　2401-2410　1999/05

DOI： 10.1093/emboj/18.9.2401 　

ISSN： 0261-4189
Ubiquitin-dependent degradation of I kappa B alpha is mediated by a ubiquitin ligase Skp1/Cul 1/F-box protein FWD1 Peer-reviewed

S Hatakeyama, M Kitagawa, K Nakayama, M Shirane, M Matsumoto, K Hattori, H Higashi, H Nakano, K Okumura, K Onoe, RA Good, K Nakayama

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA　96　(7)　3859-3863　1999/03

DOI： 10.1073/pnas.96.7.3859 　

ISSN： 0027-8424
Accelerated neutrophil apoptosis in mice lacking A1-a, a subtype of the bcl-2-related A1 gene. Peer-reviewed

Hamasaki A, Sendo F, Nakayama K, Ishida N, Negishi I, Nakayama K, Hatakeyama S

J Exp Med　188　(11)　1985-1992　1998/12/07

DOI： 10.1084/jem.188.11.1985 　
細胞周期のブレーキp27及びp57の分化と発癌に関する役割

中山敬一, 永濱裕康, 石田典子, 白根道子, 森正樹, 高橋勝彦, 畠山鎮次, 北川雅敏, 中山啓子

日本癌学会57回総会記事　20-20　1998/08
Multiple gene duplication and expression of mouse bcl-2-related genes, A1 Peer-reviewed

S Hatakeyama, A Hamasaki, Negishi, I, DY Loh, F Sendo, K Nakayama, K Nakayama

INTERNATIONAL IMMUNOLOGY　10　(5)　631-637　1998/05

DOI： 10.1093/intimm/10.5.631 　

ISSN： 0953-8178
Cip/Kip cyclin-dependent kinase inhibitors: Brakes of the cell cycle engine during development

Kei-Ichi Nakayama, Keiko Nakayama

BioEssays　20　(12)　1020-1029　1998
Publisher: John Wiley and Sons Inc.
DOI： 10.1002/(SICI)1521-1878(199812)20:12<1020::AID-BIES8>3.3.CO;2-4 　

ISSN： 0265-9247
Role of bcl-2 in the development of lymphoid cells from the hematopoietic stem cell Peer-reviewed

Y Matsuzaki, K Nakayama, K Nakayama, T Tomita, M Isoda, DY Loh, H Nakauchi

BLOOD　89　(3)　853-862　1997/02

ISSN： 0006-4971
Apoptosis during an early stage of nephrogenesis induces renal hypoplasia in bcl-2-deficient mice. Peer-reviewed

Nagata M, Nakauchi H, Nakayama K, Nakayama K, Loh D, Watanabe T

Am J Pathol　148　(5)　1601-1611　1996/05
Mice lacking p27(Kip1) display increased body size, multiple organ hyperplasia, retinal dysplasia, and pituitary tumors Peer-reviewed

K Nakayama, N Ishida, M Shirane, A Inomata, T Inoue, N Shishido, Hori, I, DY Loh, K Nakayama

CELL　85　(5)　707-720　1996/05

DOI： 10.1016/S0092-8674(00)81237-4 　

ISSN： 0092-8674
Functional significance of the Fas molecule in naive lymphocytes Peer-reviewed

S Senju, Negishi, I, N Motoyama, FP Wang, KI Nakayama, K Nakayama, PJ Lucas, S Hatakeyama, Q Zhang, S Yonehara, DY Loh

INTERNATIONAL IMMUNOLOGY　8　(3)　423-431　1996/03

DOI： 10.1093/intimm/8.3.423 　

ISSN： 0953-8178
T-B cell interaction inhibits spontaneous apoptosis of mature lymphocytes in Bcl-2-deficient mice. Peer-reviewed

Nakayama K, Nakayama K, Dustin L B, Loh D Y

J Exp Med　182　(4)　1101-1109　1995/10/01

DOI： 10.1084/jem.182.4.1101 　
ESSENTIAL ROLE FOR ZAP-70 IN BOTH POSITIVE AND NEGATIVE SELECTION OF THYMOCYTES Peer-reviewed

NEGISHI, I, N MOTOYAMA, K NAKAYAMA, K NAKAYAMA, S SENJU, S HATAKEYAMA, Q ZHANG, AC CHAN, DY LOH

NATURE　376　(6539)　435-438　1995/08

DOI： 10.1038/376435a0 　

ISSN： 0028-0836
Differential expression of bcl-2 in intestinal epithelia. Correlation with attenuation of apoptosis in colonic crypts and the incidence of colonic neoplasia. Peer-reviewed

Merritt A J, Potten C S, Watson A J, Loh D Y, Nakayama K, Nakayama K, Hickman J A

J Cell Sci　108 ( Pt 6)　2261-2271　1995/06
MASSIVE CELL-DEATH OF IMMATURE HEMATOPOIETIC-CELLS AND NEURONS IN BCL-X-DEFICIENT MICE Peer-reviewed

N MOTOYAMA, FP WANG, KA ROTH, H SAWA, K NAKAYAMA, K NAKAYAMA, NEGISHI, I, S SENJU, Q ZHANG, S FUJII, DY LOH

SCIENCE　267　(5203)　1506-1510　1995/03

DOI： 10.1126/science.7878471 　

ISSN： 0036-8075
Targeted disruption of Bcl-2 alpha beta in mice: occurrence of gray hair, polycystic kidney disease, and lymphocytopenia. Peer-reviewed

Nakayama K, Nakayama K, Negishi I, Kuida K, Sawa H, Loh D Y

Proc Natl Acad Sci U S A　91　(9)　3700-3704　1994/04/26

DOI： 10.1073/pnas.91.9.3700 　
REQUIREMENT FOR CD8 BETA-CHAIN IN POSITIVE SELECTION OF CD8-LINEAGE T-CELLS Peer-reviewed

K NAKAYAMA, K NAKAYAMA, NEGISHI, I, K KUIDA, MC LOUIE, O KANAGAWA, H NAKAUCHI, DY LOH

SCIENCE　263　(5150)　1131-1133　1994/02

DOI： 10.1126/science.8108731 　

ISSN： 0036-8075
DISAPPEARANCE OF THE LYMPHOID SYSTEM IN BCL-2 HOMOZYGOUS MUTANT CHIMERIC MICE Peer-reviewed

K NAKAYAMA, K NAKAYAMA, NEGISHI, I, K KUIDA, Y SHINKAI, MC LOUIE, LE FIELDS, PJ LUCAS, STEWART, V, FW ALT, DY LOH

SCIENCE　261　(5128)　1584-1588　1993/09

DOI： 10.1126/science.8372353 　

ISSN： 0036-8075
ACTIVATION AND REACTIVATION OF THE ATP-SENSITIVE K+ CHANNEL OF THE HEART CAN BE MODIFIED BY DRUGS Peer-reviewed

M HIRAOKA, Z FAN, T FURUKAWA, K NAKAYAMA, T SAWANOBORI

CARDIOVASCULAR DRUGS AND THERAPY　7　593-598　1993/08

ISSN： 0920-3206
AROMATIC-ALDEHYDES AND AROMATIC KETONES OPEN ATP-SENSITIVE K+ CHANNELS IN GUINEA-PIG VENTRICULAR MYOCYTES Peer-reviewed

Z FAN, K NAKAYAMA, T SAWANOBORI, M HIRAOKA

PFLUGERS ARCHIV-EUROPEAN JOURNAL OF PHYSIOLOGY　421　(5)　409-415　1992/08

ISSN： 0031-6768
Action of nicorandil on ATP-sensitive K+ channel in guinea-pig ventricular myocytes. Peer-reviewed

Nakayama K, Fan Z, Marumo F, Sawanobori T, Hiraoka M

Br J Pharmacol　103　(3)　1641-1648　1991/07

DOI： 10.1111/j.1476-5381.1991.tb09841.x 　
二次元心エコー図による左房容積の算出剖検心を用いた検討

中山啓子, 高元俊彦, 新田正男

日本臨床生理学会雑誌　21　(1)　1-9　1991/02
MULTIPLE ACTIONS OF PINACIDIL ON ADENOSINE TRIPHOSPHATE-SENSITIVE POTASSIUM CHANNELS IN GUINEA-PIG VENTRICULAR MYOCYTES Peer-reviewed

Z FAN, K NAKAYAMA, M HIRAOKA

JOURNAL OF PHYSIOLOGY-LONDON　430　273-295　1990/11

ISSN： 0022-3751
PINACIDIL ACTIVATES THE ATP-SENSITIVE K+ CHANNEL IN INSIDE-OUT AND CELL-ATTACHED PATCH MEMBRANES OF GUINEA-PIG VENTRICULAR MYOCYTES Peer-reviewed

Z FAN, K NAKAYAMA, M HIRAOKA

PFLUGERS ARCHIV-EUROPEAN JOURNAL OF PHYSIOLOGY　415　(4)　387-394　1990/01

ISSN： 0031-6768
Interrelation between pinacidil and intracellular ATP concentrations on activation of the ATP-sensitive K+ current in guinea pig ventricular myocytes Peer-reviewed

K. Nakayama, Z. Fan, F. Marumo, M. Hiraoka

Circulation Research　67　(5)　1124-1133　1990

DOI： 10.1161/01.RES.67.5.1124 　

ISSN： 0009-7330

Show all ︎Show first 5

Misc. 115

1,25(OH)₂VD₃-induced adipogenesis and osteoclastgenesis in osteblasts depend on p57^Kip2.

高橋勝彦, 難波美早, 柿崎良輔, 及川里奈, 天野均, 浦野友彦, 中山啓子, 中山敬一, 東伸昭

日本薬学会年会要旨集(Web)　144th　2024

ISSN： 0918-9823
Vitamin D₃-induced adipogenesis in osteoblasts depends on p57^Kip2

柿崎良輔, 及川里奈, 高橋優太, 天野均, 浦野友彦, 中山啓子, 中山敬一, 高橋勝彦, 東伸昭

日本薬学会年会要旨集(Web)　143rd　2023

ISSN： 0918-9823
Molecular pathogenesis in two cases of IMAGE-I syndrome with novel POLE mutations

中野智太, 森谷邦彦, 菊池敦生, 新妻秀剛, 笹原洋二, 舟山亮, 中山啓子, 城田松之, 新堀哲也, 青木洋子, 呉繁夫

日本免疫不全・自己炎症学会雑誌(Web)　2　(2)　2023

ISSN： 2435-7693
幅広い臨床症状を呈したMECOM遺伝子変異を同定した2家系

新堀哲也, 田野島玲大, 笹原洋二, 佐藤篤, 入江正寛, 南條由佳, 舟山亮, 城田松之, 阿部太紀, 奥山祐子, 石井直人, 中山啓子, 呉繁夫, 今泉益栄, 青木洋子

日本小児科学会雑誌　126　(2)　230-230　2022/02
Publisher: (公社)日本小児科学会
ISSN： 0001-6543
Methionine metabolism controls erythropoiesis by epigenetic regulation

加藤浩貴, 加藤浩貴, 加藤浩貴, NGUYEN Long, 石井悠翔, 石井悠翔, 松本光代, 三枝大輔, 舟山亮, 岡江寛明, 藤原亨, 武藤哲彦, 中山啓子, 有馬隆博, SCADDEN David, 五十嵐和彦, 張替秀郎

日本血液学会学術集会抄録(Web)　83rd　2021
多系統蛋白質症(MSP)患者の全国実態調査と診療体制構築に関する研究 HNRNPA1変異を伴う多系統蛋白質症(MSP3型)新規家系の同定

青木正志, 割田仁, 井泉瑠美子, 井泉瑠美子, 池田謙輔, 鈴木直輝, 高橋俊明, 竪山真規, 西山亜由美, 城田松之, 舟山亮, 中山啓子, 三橋里美, 西野一三, 新堀哲也, 青木洋子

多系統蛋白質症(MSP)患者の全国実態調査と診療体制構築に関する研究令和2年度総括・分担研究報告書(Web)　2021
RRAS2の活性化変異はヌーナン症候群を引き起こす

新堀哲也, 永井康貴, 藤田京志, 大橋博文, 岡本伸彦, 岡田賢, 原田敦子, 木原裕貴, THOMAS Arbogast, 舟山亮, 城田松之, 中山啓子, 阿部大紀, 井上晋一, I-CHUN Tsai, 松本直通, ERICA Davis, NICHOLAS Katsanis, 青木洋子

日本遺伝子診療学会大会プログラム・抄録集　27th　2020
満期産脳性麻痺群の網羅的遺伝学的解析 17例中9例で候補遺伝子変異を同定

竹澤祐介, 菊池敦生, 萩野谷和裕, 新堀哲也, 沼田有里佳, 松, 乾健彦, 山村菜絵子, 宮林拓矢, 安西真衣, 鈴木智, 室本, 大久保幸宗, 遠藤若葉, 冨樫紀子, 小林康子, 大沼晃, 舟山亮, 城田松之, 中山啓子, 青木洋子, 呉繁夫

日本小児科学会雑誌　123　(6)　1069-1070　2019/06
Publisher: (公社)日本小児科学会
ISSN： 0001-6543
マイクロ流体デバイスを用いた運動ニューロンの軸索病態の解析

鈴木直輝, 秋山徹也, 川田治良, 川田治良, 舟山亮, 岡野栄之, 中山啓子, 藤井輝夫, 青木正志

日本細胞生物学会大会(Web)　69回　25-25　2017/05
Publisher: (一社)日本細胞生物学会
MECOM(EVI)遺伝子変異は無巨核球性血小板減少症を伴う橈尺骨癒合症を引き起こす

新堀哲也, 内山芽里, 笹原洋二, 入江正寛, 佐藤篤, 南條由佳, 呉繁夫, 松原洋一, 今泉益栄, 青木洋子, 金子隆, 橋井佳子, 舟山亮, 長嶋剛史, 井上晋一, 中山啓子, 大薗恵一

日本小児科学会雑誌　121　(2)　300-300　2017/02
Publisher: (公社)日本小児科学会
ISSN： 0001-6543
筋ジストロフィー関連疾患の基盤的診断・治療開発研究 Dysferlinopathyおよび類似疾患の次世代シークエンサーを用いた診断および結合蛋白に注目した病態研究

青木正志, 小野洋也, 井泉瑠美子, 鈴木直輝, 菅野新一郎, 高橋俊明, 割田仁, 加藤昌昭, 西山亜由美, 島倉奈緒子, 舟山亮, 中山啓子, 新堀哲也, 青木洋子, 三宅克也, 三宅克也

筋ジストロフィー関連疾患の基盤的診断・治療開発研究平成26-28年度総括研究報告書　18‐20　2017
多系統プロテイノパチー連鎖性hnRNPA1変異に起因する孤立性封入体ミオパチー(Isolated inclusion body myopathy caused by a multisystem proteinopathy-linked hnRNPA1 mutation)

井泉瑠美子, 割田仁, 新堀哲也, 高橋俊明, 竪山真規, 鈴木直輝, 西山亜由美, 城田松之, 舟山亮, 中山啓子, 三橋里美, 西野一三, 青木洋子, 青木正志

臨床神経学　56　(Suppl.)　S280-S280　2016/12
Publisher: (一社)日本神経学会
ISSN： 0009-918X

eISSN： 1882-0654
大腸癌細胞におけるWntシグナルによるABCC3(MRP3)の発現制御

小林実, 大沼忍, 舟山亮, 井本博文, 唐澤秀明, 青木豪, 工藤克昌, 石田晶玄, 渡辺和宏, 田中直樹, 長尾宗紀, 阿部友哉, 森川孝則, 武者宏昭, 林洋毅, 中川圭, 元井冬彦, 内藤剛, 中山啓子, 海野倫明

日本外科学会定期学術集会抄録集　116回　PS-4　2016/04
Publisher: (一社)日本外科学会
小笠原諸島に侵入したグリーンアノールの進化的変化の検出と集団ゲノム解析

玉手智史, 森英章, COMPOSANO Brian J., LKRYSKO Kenneth, 舟山亮, 中山啓子, 牧野能士, 河田雅圭

日本生態学会大会講演要旨(Web)　63rd　2016
大腸癌の発癌過程においてWnt/β-cateninシグナルは抗癌剤耐性関連ABCトランスポーターであるABCC3(MRP3)の発現を抑制する

小林実, 大沼忍, 唐澤秀明, 工藤克昌, 石田晶玄, 武者宏昭, 元井冬彦, 内藤剛, 中山啓子, 海野倫明

日本癌学会総会記事　74回　J-1351　2015/10
Publisher: 日本癌学会
ISSN： 0546-0476
CRL4(VprBP) E3 Ligase Promotes Monoubiquitylation and Chromatin Binding of TET Dioxygenases (vol 342, pg 1518, 2013)

Tadashi Nakagawa, Lei Lv, Makiko Nakagawa, Yanbao Yu, Chao Yu, Ana C. D'Alessio, Keiko Nakayama, Heng-Yu Fan, Xian Chen, Yue Xiong

MOLECULAR CELL　59　(6)　2015/09

ISSN： 1097-2765

eISSN： 1097-4164
潜因性West症候群の網羅的ゲノム解析―18症例中9例で候補変異を同定―

菊池敦生, 福與なおみ, 福與なおみ, 市野井那津子, 新堀哲也, 佐藤亮, 鈴木資, 工藤宏紀, 佐藤優子, 中山東城, 柿坂庸介, 柿坂庸介, 久保田由紀, 小林朋子, 小林朋子, 舟山亮, 中山啓子, 植松貢, 青木洋子, 萩野谷和裕, 呉繁夫

日本人類遺伝学会大会プログラム・抄録集　60th　248　2015
ユビキチンリガーゼ複合体CRL4VprBPはTETタンパク質のモノユビキチン化およびクロマチンとの結合を促進する Invited

中川直, 中山啓子, Yue Xiong

ライフサイエンス新着論文レビュー "http://first.lifesciencedb.jp/archives/9768"　2015
小児科・産科領域疾患の大規模遺伝子解析ネットワークとエピゲノム解析拠点整備に関する研究神経変性疾患・遺伝性筋疾患の遺伝子解析

青木正志, 高橋俊明, 鈴木直輝, 井泉瑠美子, 加藤昌昭, 竪山真規, 割田仁, 島倉奈緒子, 安藤里紗, 舟山亮, 長嶋剛史, 中山啓子, 新堀哲也, 青木洋子

小児科・産科領域疾患の大規模遺伝子解析ネットワークとエピゲノム解析拠点整備に関する研究平成26年度委託業務成果報告書　19-20　2015
難治性筋疾患の疫学・自然歴の収集および治療開発促進を目的とした疾患レジストリー研究日本人ジスフェルリン異常症の遺伝子変異について

青木正志, 高橋俊明, 鈴木直輝, 井泉瑠美子, 加藤昌昭, 竪山真規, 割田仁, 島倉奈緒子, 安藤里紗, 舟山亮, 長嶋剛史, 中山啓子, 新堀哲也, 青木洋子

難治性筋疾患の疫学・自然歴の収集および治療開発促進を目的とした疾患レジストリー研究平成26年度総括・分担研究報告書　17‐18　2015
IDENTIFICATION OF ACQUIRED MUTATIONS BY WHOLE-GENOME SEQUENCING IN MONOMAC SYNDROME EVOLVING INTO MYELODYSPLASIA AND ACUTE LEUKEMIA

T. Fujiwara, N. Fukuhara, R. Funayama, N. Nariai, M. Kamata, T. Nagashima, K. Kojima, Y. Onishi, Y. Sasahara, K. Ishizawa, M. Nagasaki, K. Nakayama, H. Harigae

HAEMATOLOGICA　99　81-82　2014/06

ISSN： 0390-6078
眼疾患と遺伝子緑内障のゲノム解析次世代医療・個別化医療に向けて

布施昇男, 清水愛, 木村雅恵, 高野良真, 石棟, 宮澤晃子, 国松志保, 劉孟林, 渡邉亮, 安田正幸, 横山悠, 檜森紀子, 津田聡, 山本耕太郎, 中澤徹, 安田純, 勝岡史城, 小島要, 成相直樹, 松本光代, 元池育子, 長崎正朗, 木下賢吾, 五十嵐和彦, 山本雅之, 新堀哲也, 青木洋子, 松原洋一, 舟山亮, 長嶋剛史, 中山啓子, 眞島行彦, 船山智代, 田中光一, 原田高幸, 阿部春樹, 福地健郎, 安田典子, 出田秀尚, 鄭暁東, 白石敦, 大橋裕一, 石田誠夫, 原岳, 金森章泰, 山田裕子, 中村誠, 酒井寛, Richards Julia E

日本眼科学会雑誌　118　(3)　216-240　2014/03
Publisher: (公財)日本眼科学会
ISSN： 0029-0203
筋ジストロフィーおよび関連疾患の診断・治療開発を目指した基盤研究次世代シークエンサーを用いた遠位型ミオパチーにおけるゲノム解析

青木正志, 井泉瑠美子, 高橋俊明, 加藤昌昭, 鈴木直輝, 竪山真規, 割田仁, 島倉奈緒子, 安藤里紗, 舟山亮, 長嶋剛史, 中山啓子, 新堀哲也, 青木洋子

筋ジストロフィーおよび関連疾患の診断・治療開発を目指した基盤研究平成23-25年度総括研究報告書　15-16　2014
次世代シークエンサーを駆使した希少遺伝性難病の原因解明と治療法開発の研究次世代シークエンサーを用いた遺伝性筋疾患におけるゲノム解析

青木正志, 井泉瑠美子, 加藤昌昭, 割田仁, 鈴木直輝, 竪山真規, 高橋俊明, 舟山亮, 長嶋剛史, 中山啓子, 新堀哲也, 青木洋子, 松原洋一

次世代シークエンサーを駆使した希少遺伝性難病の原因解明と治療法開発の研究平成25年度総括・分担研究報告書　11-13　2014
次世代シークエンサーを駆使した希少遺伝性難病の原因解明と治療法開発の研究次世代シークエンサーを用いた遺伝性筋疾患におけるゲノム解析

青木正志, 井泉瑠美子, 加藤昌昭, 割田仁, 鈴木直輝, 竪山真規, 高橋俊明, 舟山亮, 長嶋剛史, 中山啓子, 新堀哲也, 青木洋子, 松原洋一

次世代シークエンサーを駆使した希少遺伝性難病の原因解明と治療法開発の研究平成23-25年度総合研究報告書　11-14　2014
Myofibrillar myopathyの大家系での次世代シークエンサーを用いた原因遺伝子の同定

井泉瑠美子, 新堀哲也, 青木洋子, 鈴木直輝, 加藤昌昭, 割田仁, 高橋俊明, 竪山真規, 長嶋剛史, 舟山亮, 中山啓子, 松原洋一, 青木正志

臨床神経学　53　(12)　1450-1450　2013/12
Publisher: (一社)日本神経学会
ISSN： 0009-918X

eISSN： 1882-0654
Illumina社製シークエンサーの特徴と可能性

中山啓子, 舟山亮

肝胆膵　67　(1)　25-33　2013/07/28
Publisher: 株式会社アークメディア
Illumina社製シークエンサーの特徴と可能性

中山啓子, 舟山亮

肝胆膵　67　(1)　1-164　2013/07/28
Publisher: アークメディア
次世代シークエンサーを用いた新規発達緑内障遺伝子の探索

清水愛, 新堀哲也, 青木洋子, 松原洋一, 舟山亮, 長嶋剛史, 中山啓子, 渡邉亮, 長崎正朗, 布施昇男, 中澤徹

日本眼科学会雑誌　117　2013

ISSN： 0029-0203
Myofibrillar myopathyの大家系での次世代シークエンサーを用いた原因遺伝子の同定

井泉瑠美子, 井泉瑠美子, 新堀哲也, 青木洋子, 鈴木直輝, 加藤昌昭, 割田仁, 高橋俊明, 竪山真規, 長嶋剛史, 舟山亮, 中山啓子, 松原洋一, 青木正志

日本神経学会学術大会プログラム・抄録集　54th　332　2013
Myofibrillar myopathyの大家系における次世代型シークエンサーを用いた新たな原因遺伝子の同定

井泉瑠美子, 新堀哲也, 青木洋子, 鈴木直輝, 加藤昌昭, 割田仁, 高橋俊明, 竪山真規, 長嶋剛史, 舟山亮, 阿部康二, 中山啓子, 青木正志, 松原洋一

日本人類遺伝学会大会プログラム・抄録集　58th　136　2013
Spontaneous Development of Arrhythmogenic Right Ventricular Cardiomyopathy in Mice Overexpressing Dominant-Negative Rho-Kinase in Vascular Smooth Muscle Cells

Alia Ellawindy, Kimio Satoh, Shin-ichi Tanaka, Shohei Ikeda, Toru Shimizu, Kazuki Noda, Yoshihiro Fukumoto, Kazuto Kobayashi, Keiko Nakayama, Hiroaki Shimokawa

CIRCULATION　126　(21)　2012/11

ISSN： 0009-7322

eISSN： 1524-4539
Close Up実験法(Series 224) 高感度に多型を検出するためのエキソーム・シークエンシング (実験医学)

舟山亮, 長嶋剛史, 中山啓子

実験医学　30　(4)　629-634　2012/03
Bach1によるPparγ遺伝子発現抑制を介した脂肪細胞分化阻害

松本光代, 松本光代, 白木琢磨, BURYDUN Andrey, 舟山亮, 西田有一郎, 中山啓子, 八重樫伸生, 五十嵐和彦

日本分子生物学会年会プログラム・要旨集(Web)　35th　2012
Bach1の転写制御標的遺伝子の同定

松本光代, 松本光代, BRYDUN Andrey, 中目亜矢子, 舟山亮, 西田有一郎, 中山啓子, 八重樫伸生, 五十嵐和彦

日本分子生物学会年会プログラム・要旨集(Web)　34th　2011
マウス胎仔線維芽細胞におけるFbxw 7による細胞周期抑制因子の制御 (日本外科学会雑誌)

益田邦洋, 石川善則, 山本久仁治, 片寄友, 小野山一郎, 中山敬一, 海野倫明, 中山啓子

日本外科学会雑誌　111　(臨増2)　465-465　2010/03
細胞周期を抑制するユビキチンリガーゼと発癌 (炎症と免疫)

中山啓子

炎症と免疫　17　(2)　29-34　2009/02/20
がん抑制遺伝子変異による組織特異的発がん機構の解明 (癌と人)

中山啓子

癌と人　35　44-45　2008/04/25
SCF型ユビキチンリガーゼによる細胞周期と発癌 (実験医学)

中山敬一, 中山啓子

実験医学　26　166-174　2008/02/01
p27ノックアウトマウスにおけるカイニン酸誘発応答の増大 (日本薬学会年会要旨集)

赤芝洋紀, 上山千紘, 中山啓子, 中山敬一, 西山信好, 松木則夫

日本薬学会年会要旨集　127年会　(2)　126-126　2007/03
【シグナル伝達病を知るその分子機序解明から新たな治療戦略まで】基礎編細胞増殖・分化・死のシグナル伝達細胞周期とシグナル伝達 (遺伝子医学MOOK)

中山啓子

遺伝子医学MOOK　(6)　101-106　2006/12
ユビキチンリガーゼコンポーネントFbw7は造血幹細胞の静止状態を制御する (臨床血液)

松岡佐保子, 尾池雄一, 小野山一郎, 田久保圭誉, 伊藤圭介, 新井文用, 岩間厚志, 池田康夫, 中山啓子, 中山敬一, 須田年生

臨床血液　47　(9)　1035-1035　2006/09
Publisher: (一社)日本血液学会-東京事務局
ISSN： 0485-1439

eISSN： 1882-0824
肥満・糖尿病発症におけるユビキチンリガーゼSkp2の役割 (肥満研究)

酒井太門, 阪上浩, 中村武寛, 岡田潮, 永礼智基, 春日雅人, 松木泰, 渡辺英二郎, 平松隆司, 中山啓子

肥満研究　12　(Suppl.)　178-178　2006/09
【ユビキチン-プロテアソーム系とオートファジー作動機構と病態生理】ユビキチン系の生理と病態ユビキチンと細胞周囲制御・癌 G1期からS期を制御するユビキチン系 (蛋白質・核酸・酵素)

中山敬一, 中山啓子

蛋白質・核酸・酵素　51　(10)　1362-1369　2006/08
Publisher: 共立出版
ISSN： 0039-9450
Ubiquitin ligases: cell-cycle control and cancer

KI Nakayama, K Nakayama

NATURE REVIEWS CANCER　6　(5)　369-381　2006/05

DOI： 10.1038/nrc1881 　

ISSN： 1474-175X
肥満・糖尿病発症におけるユビキチンリガーゼSkp2の役割 (糖尿病)

酒井太門, 阪上浩, 中村武寛, 岡田潮, 岡田裕子, 高島康弘, 永礼智基, 中村恭子, 春日雅人, 中山啓子

糖尿病　49　(Suppl.1)　S267-S267　2006/04
p27の増加はDNA損傷後の中心体数異常の抑制に必要である

杉原英志, 豊島秀男, 中山啓子, 中山敬一, 佐谷秀行, 三輪正直

日本癌学会学術総会記事　65th　2006
Skp2による脂肪細胞数制御機構の肥満発症における役割 (肥満研究)

酒井太門, 中山啓子

肥満研究　11　(Suppl.)　154-154　2005/09
【細胞周期の最前線明らかになるその制御機構複製のライセンス化,チェックポイント,分裂期キナーゼなど精緻な分子機構と生命現象とのかかわり】 G2期からM期へこのダイナミックな変化の分子メカニズム p27の分解はM期の進行に必要である (実験医学)

中山啓子

実験医学　23　(9)　1383-1388　2005/05
p27はDNAダメージ後の中心体数異常を抑制する

杉原英志, 杉原英志, 斎藤総一郎, 斎藤総一郎, 豊島秀男, 中山啓子, 中山敬一, 佐谷秀行, 三輪正直

日本分子生物学会年会講演要旨集　28th　2005
p27はDNA損傷後の中心体過剰複製を抑制する

杉原英志, 斎藤総一郎, 新田孝幸, 豊島秀男, 中山啓子, 中山敬一, 佐谷秀行, 三輪正直

日本癌学会学術総会記事　64th　2005
CDKインヒビターの生理的機能 (生化学)

中山啓子

生化学　76　(9)　1191-1202　2004/09
細胞周期とがん M期進行におけるp27分解の重要性 (Cancer Science)

中山啓子, 三宅智, 中山敬一

Cancer Science　95　(Suppl.)　63-63　2004/09
EIDファミリータンパクは転写抑制によって分化を阻害する (Cancer Science)

三宅智, 中山啓子

Cancer Science　95　(Suppl.)　44-44　2004/09
脂肪細胞の数とサイズを決定する細胞周期制御機構の糖代謝における重要性 (Diabetes Frontier)

中村武寛, 阪上浩, 酒井太門, 岡田潮, 岡田裕子, 高島康弘, 森要之, 中村恭子, 小川渉, 中山敬一, 中山啓子, 春日雅人

Diabetes Frontier　15　(4)　574-575　2004/08
細胞周期と再生 p27ブレーキの分解による制御機構 (炎症・再生)

中山敬一, 中山啓子

炎症・再生　24　(4)　369-369　2004/07
脂肪細胞の数とサイズを決定する細胞周期制御機構の糖代謝における重要性 1 (日本内分泌学会雑誌)

中村武寛, 阪上浩, 酒井太門, 岡田潮, 岡田裕子, 高島康弘, 森要之, 中村恭子, 小川渉, 中山啓子, 中山敬一, 春日雅人

日本内分泌学会雑誌　80　(1)　106-106　2004/04
脂肪細胞の数とサイズを決定する細胞周期制御機構の糖代謝における重要性 2 (糖尿病)

中村武寛, 阪上浩, 小川渉, 酒井太門, 岡田潮, 森要之, 岡田裕子, 高島康弘, 中村恭子, 春日雅人, 中山敬一, 中山啓子

糖尿病　47　(Suppl.1)　S101-S101　2004/04
【蛋白質のユビキチン化と疾患】生命現象とユビキチン化時計遺伝子産物のユビキチン化による分解機構 (現代医療)

中山啓子

現代医療　36　(4)　893-898　2004/04
Publisher: 現代医療社
ISSN： 0533-7259
B細胞へのトレランス誘導とPKC-δ (臨床免疫)

宮本顕友, 中山啓子, 中山敬一

臨床免疫　39　(2)　236-240　2003/02
Publisher: 科学評論社
ISSN： 0386-9695
【次々と解明されるユビキチンの多彩な役割】細胞周期を制御する二大ユビキチンリガーゼ APC/CとSCF複合体 (実験医学)

中山敬一, 中山啓子

実験医学　21　(3)　358-364　2003/02
自己免疫疾患とPKC-δ (感染・炎症・免疫)

宮本顕友, 中山啓子, 中山敬一

感染・炎症・免疫　32　(4)　302-303　2002/12
Bリンパ球の増殖を抑制するシグナル伝達分子PKC-δ (実験医学)

中山敬一, 宮本顕友, 中山啓子

実験医学　20　1438-1441　2002/07
PKC-δノックアウトマウスにおけるB細胞の過剰増殖と自己免疫疾患 (細胞工学)

中山啓子, 宮本顕友, 中山敬一

細胞工学　21　(7)　778-779　2002/06
【細胞周期サイクリンの発見から20年解明が進む制御機構と発癌メカニズム】 CDKインヒビターp27Kip1のタンパク質分解による制御機構 (実験医学)

中山啓子, 中山敬一

実験医学　20　(4)　526-531　2002/03
【集まれ!モデル生物たち】ヒトに最も近いモデル生物-マウス (細胞工学)

中山啓子, 中山敬一

細胞工学　21　(1)　31-36　2001/12
細胞周期の開始と停止の制御機構静止期から増殖期へ p27分解に関わる二つのユビキチンリガーゼ (生化学)

中山敬一, 嘉村巧, 原太一, 畠山鎮次, 中山啓子

生化学　73　(8)　636-636　2001/08
Regulation of the cell cycle at the G(1)-S transition by proteolysis of cyclin E and p27(Kip1)

K Nakayama, S Hatakeyama, K Nakayama

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS　282　(4)　853-860　2001/04

DOI： 10.1006/bbrc.2001.4627 　

ISSN： 0006-291X
【タンパク質分解の最前線2001】ユビキチンシステムユビキチンリガーゼ研究の最前線 SCF複合体の多面的な作用機構 (実験医学)

中山敬一, 中山啓子

実験医学　19　(2)　132-141　2001/01
GABAA受容体情報伝達におけるp130,新規IP3結合蛋白質の役割 (生化学)

兼松隆, 吉村研治, 鍋倉淳一, 赤池紀生, 中山啓子, 中山敬一, 平田雅人

生化学　72　(8)　990-990　2000/08
蛋白質分解制御による細胞分裂周期の調節サイクリンE及びp27Kip1のユビキチン化因子のノックアウトマウスによる機能解析 (生化学)

中山敬一, 畠山鎮次, 北川雅敏, 小南欽一郎, 中山啓子

生化学　72　(8)　596-596　2000/08
【拡大するユビキチンの世界】 SCF複合体千の顔をもつユビキチンリガーゼ (実験医学)

中山敬一, 中山啓子

実験医学　18　(11)　1457-1464　2000/07
SCF複合体によるユビキチン依存性のタンパク質分解 (生化学)

中山啓子, 中山敬一

生化学　72　(2)　113-117　2000/02
Publisher: 日本生化学会
ISSN： 0037-1017
細胞周期制御因子およびシグナル伝達因子の分解の分子機構 (生化学)

北川雅敏, 畠山鎮次, 白根道子, 中山啓子, 中山敬一

生化学　71　(8)　1042-1042　1999/08
NFκB活性化シグナルと生命現象 IκBのユビキチンリガーゼSCFFWD1複合体の同定 (生化学)

中山敬一, 畠山鎮次, 北川雅敏, 白根道子, 松本雅記, 服部公彦, 中山啓子

生化学　71　(8)　686-686　1999/08
Chk1キナーゼ欠損マウスを用いたマウスChk1キナーゼの機能解析

高井裕之, 冨永薫, 本山昇, 池田恭治, 中西真, 永浜裕康, 中山啓子, 中山敬一

日本分子生物学会年会プログラム・講演要旨集　22nd　1999
Genomic DNA cloning and analysis of a novel F-box protein, FWD2

MIURA Masatoku, HATAKEYAMA Shigetsugu, NAKAYAMA Keiko, KITAGAWA Masatoshi, NAKAYAMA Kei-ichi

21　459-459　1998/12/01
Biochemical analysis of binding motif between murine Skp1 and F-box protein, FWD1

HATTORI Kimihiko, HATAKEYAMA Shigetsugu, NAKAYAMA Keiko, KITAGAWA Masatoshi, NAKAYAMA Kei-ichi

21　460-460　1998/12/01
Molecular cloning and cell biological analysis of mouse novel F-box protein cDNAs

HATAKEYAMA Shigetsugu, KITAGAWA Masatoshi, SHIRANE Michiko, MATSUMOTO Masaki, HATTORI Kimihiko, NAKAYAMA Keiko, NAKAYAMA Kei-ichi

21　460-460　1998/12/01
Down-regulation of p27^<Kip1> by two mechanisms, ubiquitin-mediated degradation and site-specific cleavage

SHIRANE Michiko, HARUMIYA Yumiko, ISHIDA Noriko, HATAKEYAMA Shigetsugu, NAKAYAMA Keiko, NAKAYAMA Kei-ichi, KITAGAWA Masatoshi

21　457-457　1998/12/01
Molecular Cloning and biochemical analysis of mouse ubiquitin conjugating enzyme, Ubc M5

YAMANAKA Atsushi, HATAKEYAMA Shigetsugu, KITAGAWA Masatoshi, NAKAYAMA Keiko, NAKAYAMA Kei-ichi

21　500-500　1998/12/01
Molecular cloning and analysis of murine Skp2 cDNA and genomic DNA

MATSUMOTO Masatoshi, HATAKEYAMA Shigetsugu, KITAGAWA Kyouko, KITAGAWA Masatoshi, NAKAYAMA Keiko, NAKAYAMA Kei-ichi

21　500-500　1998/12/01
Generation and analysis of thymocyte specific expressed p27^<Kip1> transgenic mice

TSUKIYAMA Tadasuke, HATAKEYAMA Shigetsugu, KITAGAWA Masatoshi, NAKAYAMA Keiko, NAKAYAMA Kei-ichi

21　589-589　1998/12/01
Degradation mechanizms of p27^<kip1> and cell -growth related proteins

KITAGAWA Masatoshi, HATAKEYAMA Shigetsugu, SHIRANE Michiko, ISHIDA Noriko, HIRAI Aizan, TANAKA Tomoaki, NAKAYAMA Keiko, NAKAYAMA Kei-ichi

日本分子生物学会年会プログラム・講演要旨集　21　217-217　1998/12/01
マウスSkp1 cDNA及びgenomic DNAのクローニング

南嶋洋司, 畠山鎮次, 豊野孝, 北川雅敏, 中山啓子, 中山敬一

日本分子生物学会年会プログラム・講演要旨集　21st　460　1998/11
細胞周期関連遺伝子 (臨床免疫)

中山啓子, 中山敬一

臨床免疫　30　(4)　497-504　1998/04
Bcl-2ファミリーによる生体制御 (現代化学(増刊）)

中山敬一, 中山啓子

現代化学(増刊）　35　87-95　1997/12/05
Bcl-2ファミリ-による生体制御 (アポト-シス研究の新展開) -- (アポト-シスによる生体制御)

中山敬一, 中山啓子

現代化学増刊　(35)　87-95　1997/12
Publisher: 東京化学同人
ISSN： 0910-4747
CDKインヒビター欠損マウスにおける病態 (臨床免疫)

中山啓子, 中山敬一

臨床免疫　29　(10)　1285-1291　1997/10
サイトカインと免疫調節 p27Kip1によるT細胞の増殖制御 (Molecular Medicine)

中山啓子, 中山敬一

Molecular Medicine　34　(臨増免疫1997-98)　80-88　1997/09
免疫系の発生と分化 T細胞の発生と分化 T細胞分化における細胞周期調節 (実験医学)

中山啓子, 中山敬一

実験医学　15　(11)　1315-1321　1997/07
ゲルの写真にこだわる (実験医学)

中山敬一, 中山啓子

実験医学　15　932-933　1997/06
平底チューブを使いこなそう (実験医学)

中山敬一, 中山啓子

実験医学　15　568-569　1997/04
迅速Mini-prep(Quick-prep)法 (実験医学)

中山敬一, 中山啓子

実験医学　15　(2)　185-186　1997/02
Regulation of Cell Growth by p27^Kip1 and p57^Kip2 and Their involvement in Carcinogenesis.

中山敬一, 森正樹, 高橋勝彦, 中山啓子

日本分子生物学会年会プログラム・講演要旨集　20th　1997
Creation and analysis of p57^KIP2 knock-out mice.

高橋勝彦, 中山啓子, 中山敬一, 松村義裕, 高橋実, 佐藤正伸, 立石裕, 根岸泉, LOH D Y

日本分子生物学会年会プログラム・講演要旨集　20th　1997
p27Kip1ノックアウトマウス細胞周期抑制分子の生理的役割 (実験医学)

中山敬一, 中山啓子

実験医学　15　(1)　88-90　1997/01
細胞周期調節のT細胞分化に与える影響 : p27ノックアウトマウスを用いた解析

中山啓子, 石田典子, 白根道子, 猪又晃, 井上智彰, 宍戸信之, 堀井郁夫, LOH Dennis Y., 中山敬一

日本分子生物学会年会プログラム・講演要旨集　19　42-42　1996/08
p27ノックアウトマウスにおける成長の促進と多臓器腫大

中山啓子, 石田典子, 白根道子, 猪又晃, 井上智彰, 宍戸信之, 堀井郁夫, LOH Dennis Y., 中山敬一

日本分子生物学会年会プログラム・講演要旨集　19　646-646　1996/08/01
bcl-2遺伝子ファミリーとアポトーシス Bcl-2欠損マウス (血液・腫瘍科)

中山敬一, 中山啓子

血液・腫瘍科　32　(4)　302-311　1996/04
Role of ZAP-70 in T cell differentiation.

根岸泉, 本山昇, 中山敬一, 中山啓子, 千住覚, 森真弥, LOH D Y

日本免疫学会総会・学術集会記録　26　1996

ISSN： 0919-1984
T細胞分化とアポトーシス (実験医学)

中山敬一, 中山啓子

実験医学　13　(16)　1952-1959　1995/10
遺伝子操作による免疫異常モデル CD4およびCD8欠損マウス (炎症と免疫)

中山啓子, 中山敬一

炎症と免疫　3　(6)　642-650　1995/10
T細胞の選択とアポトーシス (Mebio)

中山敬一, 中山啓子

Mebio　12　(10)　79-86　1995/10
T細胞の分化と表面機能分子 T細胞の分化とCD8β鎖ノックアウトマウスによる解析 (臨床免疫)

中山啓子, 中山敬一

臨床免疫　27　(8)　926-937　1995/08
Publisher: 科学評論社
ISSN： 0386-9695
リンパ球の細胞死とBcl-2 (臨床免疫)

中山敬一, 中山啓子

臨床免疫　27　(7)　829-838　1995/07
Publisher: 科学評論社
ISSN： 0386-9695
リンパ球のシグナル伝達 Bcl-2によるリンパ球細胞死の調節 (Molecular Medicine)

中山敬一, 中山啓子

Molecular Medicine　32　(臨増)　52-63　1995/06
Publisher: 中山書店
ISSN： 0918-6557
ZAP-70欠損マウスにおけるT細胞分化

根岸泉, 本山昇, 中山敬一, 中山啓子, 千住覚, 畠山鎮次, ZHAN Q, CHAN A C, LOH D Y

日本免疫学会総会・学術集会記録　25　1995

ISSN： 0919-1984
Bcl-2欠損マウスにおけるリンパ球の消失 (臨床免疫)

中山敬一, 中山啓子

臨床免疫　26　(10)　1154-1162　1994/10
Bcl-2ノックアウトマウス (免疫Immunology Frontier)

中山敬一, 中山啓子

免疫Immunology Frontier　4　(3)　175-182　1994/06
Bcl-2欠損マウスダブルノックアウト法を用いて (実験医学)

中山敬一, 中山啓子

実験医学　12　(7)　851-854　1994/05
Life-span of immature lymphocytes but not mature lymphocytes is shortened in bcl-x deficient chimeric mice.

本山昇, WANG F, 中山敬一, 中山啓子, 根岸泉, 千住悟, ZHANG Q, LOH D Y

日本免疫学会総会・学術集会記録　24　1994

ISSN： 0919-1984
免疫細胞の分化遺伝子ターゲティングによる展開 Bcl-2欠損マウスダブルノックアウト法を用いて (Medical Immunology)

中山敬一, 中山啓子

Medical Immunology　27　(1)　73-82　1994/01
Pinacidilの心筋ATP感受性Kチャンネルに対する作用 (最新醫學)

中山啓子, 范錚, 丸茂文昭, 平岡昌和

最新醫學　45　2470-2473　1990/12
K+ channel開口薬,NicorandilとPinacidilの心筋ATP感受性K+ channelへの作用とその差異 (Therapeutic Research)

平岡昌和, 范錚, 中山啓子

Therapeutic Research　11　(5)　1374-1375　1990/05

Show all ︎Show first 5

Books and Other Publications 8

細胞工学

監修中山敬一, 学生体防御医学研究所, ed. 中山啓子

株式会社学研メディカル秀潤社　2012/05
The Cell Cycle Principles of Control

メディカル・サイエンス, インターナショナル ed

メディカル・サイエンス・インターナショナル　2008/05/30
キーワードで理解する細胞周期イラストマップ

中山敬一, 中山啓子

羊土社　2005/02/25
発生における細胞増殖制御

竹内隆, 岸本健雄, 中山啓子, 中山敬一

Springer　2004/02/13
わかる実験医学シリーズ「細胞周期がわかる」

中山敬一, 中山啓子, 中山敬一

羊土社　2001/02/15
わかる細胞周期と癌

田矢洋一, 中山啓子, 中山敬一

羊土社　2000/04
新用語ライブラリー細胞周期

田矢洋一, 野島博, 花岡文雄, 中山敬一, 中山啓子

羊土社　1999/08
Molecular Medicine Vol.34 臨床増刊号ノックアウトマウス・データブック

黒川清, 笹月健彦, ds, 中山啓子, 中山敬一

中山書店　1997/12/25

Show all Show first 5

Presentations 140

がん細胞における翻訳制御因子 EEF1A アイソフォームの解析

築地巧人, 中山啓子

第38回日本癌学会　2021/10/01
RBFOX2 と PTBP1 は大腸癌細胞において PTK2 マイクロエクソンのスプライシングを制御する

大平優丈, 中山啓子

第38回日本癌学会　2021/10/02
RNAによるALS関連因子TDP-43凝集体の形成と維持

中川直, 細井徹, 中山啓子

第141日本薬学会年会
VCP阻害薬によるALS関連因子TDP-43の凝集体形成抑制

中川直, 余玉嬌, 細井徹, 中山啓子

第94回日本薬理学会年会　2021/03/09
An Amyotrophic Lateral Sclerosis-Associated Mutant of C21ORF2 Is Stabilized by NEK1-Mediated Hyperphosphorylation and the Inability to Bind FBXO3

Tadashi Nakagawa, Yasuaki Watanabe, Naoki Suzuki, Hitoshi Warita, Masashi Aoki, Keiko Nakayama

31ST INTERNATIONAL SYMPOSIUM ON ALS/MND　2020/12/11
自閉症関連タンパク質USP9XによるSETD5を介したrDNAのエピジェネティック制御

中川直, 中川真喜子, 細井徹, 中山啓子

第43回日本分子生物学会年会　2020/12/02
蛋白分解異常による新規筋萎縮性側索硬化症の発症機構

渡辺靖彰、中川直、鈴木直輝、割田仁、中山啓子、青木正志

第42回日本神経科学大会　2019/07/25
ユビキチン化によるヒストンシャペロンFACT複合体の機能制御 Invited

中川直、中山啓子

第41回日本分子生物学会　2018/11/29
Cyclin Fの突然変異はVCPのATPase活性に影響することによりALS発症に関与する可能性がある

余玉嬌、諸星茜、中川直、中山啓子

第41回日本分子生物学会　2018/11/29
ユビキチン化によるヒストンシャペロンFACTの制御

中川直、諸星茜、中山啓子

東北大学知のフォーラム5周年記念特別講演会　2018/07/29
ALS患者で同定されたC21ORF2変異によるNEK1の制御

渡辺靖彰、中川直、鈴木直輝、割田仁、中山啓子、青木正志

第59回日本神経学会総会　2018/05
ユビキチンリガーゼβ-TrCPによるオス生殖細胞の減数分裂開始制御

中川直、中山啓子

第40回日本分子生物学会　2017/12/08
VCPはALSに関連したTDP-43の細胞質凝集体形成に影響する

余玉嬌、中川直、中山啓子

第40回日本分子生物学会　2017/12/07
ユビキチンリガーゼβ-TrCPによるオス生殖細胞の減数分裂開始制御

中川直、中野星児、久志瞭、中山啓子

先端モデル動物支援プラットフォーム若手技術支援講習会　2017/09/08
Molecular analysis of Amyotrophic Lateral Sclerosis associated Cyclin F mutants

Yujiao Yu,Akane Morohoshi,Tadashi Nakagawa,Keiko Nakayama

第39回日本分子生物学会年会　2016/12/01
ユビキチン化によるFACT複合体の機能制御

諸星茜、中川直、中山啓子

第39回日本分子生物学会年会　2016/12/01
H3K27ヒストン修飾によるエンハンサー制御のゲノムワイド解析

細金正樹、中山啓子

第39回日本分子生物学会年会　2016/12/01
大腸がんにおけるタンパク質シトルリン化酵素PADI2の役割

舟山亮、谷口肇、水間正道、藤島史喜、小林実、大沼忍、海野倫明、中山啓子

第39回日本分子生物学会年会　2016/11/30
Reactivation of cell proliferation by continuous TGF-β treatment

Tadashi Nakagawa,Masaki Hosogane,Ryo Funayama,Keiko Nakayama.

第39回日本分子生物学会年会　2016/11/30
セルトリ細胞特異的β-TrCPノックアウトマウスの解析

諸星茜, 中川直, 中野星児

第38回日本分子生物学会年会　2015/12/01
がん原遺伝子RASによる遺伝子サイレンシングの分子機構の解析

舟山亮, 細金正樹, 長嶋剛史

第38回日本分子生物学会年会　2015/12/01
TGF-β刺激によるTFIID構成因子TAF7の分解とその役割の解明

中川直, 細金正樹, 舟山亮

第38回日本分子生物学会年会　2015/12/01
マウスES細胞におけるgemininの細胞周期と分化に対する制御

福本恵美子, Kundu, Lena Rani, 佐藤聡一郎, 細金正樹, 中山啓子

第37回日本分子生物学会年会　2014/11/25
H3K27me3修飾パターン形成過程の時系列解析

細金正樹, 舟山亮, 城田松之, 中山啓子

第37回日本分子生物学会年会　2014/11/25
精子形成におけるE3ユビキチンリガーゼβ-TrCPの新規基質の同定と解析

久志瞭, 中川直, 中野星児, 遠藤尚博, 中山啓子

第37回日本分子生物学会年会　2014/11/25
TFIID complex changes accompany epithelial-mesenchymal transition (EMT)

Nakagawa, Tadashi, Nakayama, Keiko

第37回日本分子生物学会年会　2014/11/25
がん遺伝子による転写変化とエピゲノム変化

第23回長崎障害者支援再生医療研究会　2014/11/04
H3K27メチル化に先立って転写は抑制される

新学術領域研究細胞運命制御第５回領域会議　2014/09/03
転写制御とヒストン修飾

順天堂大学最先端・次世代シンポジウム　2013/11/30
Functional Analysis of F-box protein in vivo

第３５回内藤コンファレンス　2013/07/09
Change of Trimethylation of H3K27 Occurs after Ras-mediated Transcriptional Regulation International-presentation

NIH-Tohoku University-JSPS Symposium　2013/05/09
RNFによるKu80ユビキチン化を介したNHEJ修復制御

石田典子, 家村俊一郎, 安井明, 夏目徹, 中山啓子

第35回日本分子生物学会年会　2012/12/14
がん遺伝子Rasによる転写制御後にH3K27me3修飾変化が誘起される

細金正樹, 舟山亮, 西田有一郎, 長嶋剛史, 中山啓子

第35回日本分子生物学会年会　2012/12/14
がん遺伝子Rasによる転写制御後にH3K27me3修飾変化が誘起される

細金正樹, 舟山亮, 西田有一郎, 長嶋剛史, 中山啓子

第35回日本分子生物学会年会　2012/12/14
shRNAライブラリのスクリーニングによる遺伝子サイレンシング機構の解析

舟山亮, 細金正樹, 長嶋剛史, 中山啓子

第35回日本分子生物学会年会　2012/12/13
mES細胞におけるGemininの細胞周期‐ 細胞分化に果たす役割

Kundu Lena R, 中山啓子

第35回日本分子生物学会年会　2012/12/13
Nrf2-小Maf転写因子のゲノムワイド解析による遺伝子発現制御システムの解明

弘津陽介, 勝岡史城, 舟山亮, 長嶋剛史, 西田有一郎, 中山啓子, ジェームスダグラスエンゲル, 山本雅之

第35回日本分子生物学会年会　2012/12/13
Fbxw7はマウスの肝臓において脂質代謝及び細胞分化決定を制御する

磯下理恵子, 小野山一郎, 鈴木淳史, 松本有樹修, 冨田謙吾, 片桐秀樹, 尾池雄一, 中山啓子, 中山敬一

第35回日本分子生物学会年会　2012/12/13
mES細胞におけるGemininの細胞周期‐ 細胞分化に果たす役割

Kundu Lena R, 中山啓子

第35回日本分子生物学会年会　2012/12/13
RASシグナルによる選択的スプライシング変化の網羅的解析

松島和洋, 舟山亮, 長嶋剛史, 中山啓子

第35回日本分子生物学会年会　2012/12/12
Bach1によるPparγ遺伝子発現抑制を介した脂肪細胞分化阻害

松本光代, 白木琢磨, ブリドンアンドレイ, 舟山亮, 西田有一郎, 中山啓子, 八重樫伸生, 五十嵐和彦

第35回日本分子生物学会年会　2012/12/11
Change of Trimethylation of H3K27 Occurs after Ras-mediated Transcriptional Regulation International-presentation

Keiko Nakayama

CSI-GCOE Joint Workshop on Genome Science　2012/08/21
Analysis of Geminin-deficient mES cells International-presentation

Kundu, L. R, Nakayama, K

2012 Cold Spring Harbor Laboratory on the Cell Cycle　2012/05/15
Exome sequencing of patients with autophagic vacuolar myopathies

Funayama, R, Nagashima, T, Nakayama, K

Winter Camp of GCOE 2012 〜Challenge to Advanced Research through Interdisciplinary Exchange 〜 Tohoku University Global COE for Conquest of Signal Transduction Diseases with “Network Medicine”　2012/02/04
Geminin deletion in hematopoietic stem cells promotes differentiation of megakaryocytes and platelets

Nakano, S, Sasaki, Y, Motohashi, H, Nakayama, K

Winter Camp of GCOE 2012 〜Challenge to Advanced Research through Interdisciplinary Exchange 〜 Tohoku University Global COE for Conquest of Signal Transduction Diseases with “Network Medicine”　2012/02/04
Change of trimethylation of H3K27 occurs after Ras-mediated transcriptional regulation

Hosogane, M, Funayama, R, Nishida, Y, Nagashima, T, Nakayama, K

Winter Camp of GCOE 2012 〜Challenge to Advanced Research through Interdisciplinary Exchange 〜 Tohoku University Global COE for Conquest of Signal Transduction Diseases with “Network Medicine”　2012/02/04
Comprehensive analysis of altemative splicing by next-generation sequencer

Matsushima, W, Funayama, R, Nagashima, T, Nakayama, K

Winter Camp of GCOE 2012 〜Challenge to Advanced Research through Interdisciplinary Exchange 〜 Tohoku University Global COE for Conquest of Signal Transduction Diseases with “Network Medicine”　2012/02/04
Selective epigenetic gene regulation in primordial germ cells

Mochizuki, K, Hosogane, M, Nakayama, K, Matsui, Y

Winter Camp of GCOE 2012 〜Challenge to Advanced Research through Interdisciplinary Exchange 〜 Tohoku University Global COE for Conquest of Signal Transduction Diseases with “Network Medicine”　2012/02/04
胆道癌における癌関連遺伝子の変異解析

高野成尚, 中山啓子, 海野倫明

第５回リトリート大学院生研究発表会　2012/01/21
がん遺伝子Rasによる転写制御後にH3K27me3修飾変化が誘起される

細金正樹, 舟山亮, 西田有一郎, 長嶋剛史, 中山啓子

第５回リトリート大学院生研究発表会　2012/01/21
Ku80のユビキチン化を介したＤＮＡ二重鎖切断修復制御

石田典子, 家村俊一郎, 安井明, 夏目徹, 中山啓子

第３４回日本分子生物学会年会　2011/12/13
Ku80のユビキチン化を介したＤＮＡ二重鎖切断修復制御

石田典子, 家村俊一郎, 安井明, 夏目徹, 中山啓子

第３４回日本分子生物学会年会　2011/12/13
Change of trimethylation of H3K27 occurs after Ras-mediated transcriptional regulation

Hosogane, M, Funayama, R, Nishida, Y, Nagashima, T, Nakayama, k

第３４回日本分子生物学会年会　2011/12/13
Change of trimethylation of H3K27 occurs after Ras-mediated transcriptional regulation

Hosogane, M, Funayama, R, Nishida, Y, Nagashima, T, Nakayama, k

第３４回日本分子生物学会年会　2011/12/13
Bach1の転写制御標的遺伝子の同定

松本光代, ブリドンアンドレイ, 中目亜矢子, 舟山亮, 西田有一郎, 中山啓子, 八重樫伸生, 五十嵐和彦

第３４回日本分子生物学会年会　2011/12/13
Cdt1 inhibitor, Geminin negatively regulates thrombopoiesis

Nakayama, K

The 5th International Workshop on Cell Regulations in Division and Arrest　2011/10/23
Changes of trimethylation of H3K27 occurs after Ras-mediated transcriptional regulation

Hosogane, M, Funayama, R, Nishida, Y, Nagashima, T, Nakayama, K

The 5th International Workshop on Cell Regulations in Division and Arrest　2011/10/23
H3K27 is tri-methylated after Ras-mediated regional silencing

Hosogane, M, Nakayama, K

70th Annual Meeting of the Japanese Cancer Association　2011/10/03
Ku80のユビキチン化を介したDNA二重鎖切断修復制御

中山啓子, 石田典子

第１７回国際癌治療増感研究会　2011/06/24
Tissue-specific fnction of Fbxw7 revealed by conditional gene targeting in multiple mouse organs International-presentation

Nakayama,K, Onoyama,I, Matsumoto,A, Ishikawa,T, Aoyama,S, Nakayama,K

Cold Spring Harbor Laboratory 2011Meeting & Courses　2011/05/17
Ras-mediated regional silencing around Fas locus.

Masaki Hosogane, Ryo Funayama, Yuichiro Nishida, Keiko Nakayama

東北大学グローバルCOE Network Medicine創生拠点冬の合宿2011　2011/02/05
Geminin deletion in hematopeietic stem cells promotes differentiation of megakaryocytes and platelets.

Seiji Nakano, Yousuke Sasaki, Hozumi Motohashi, Keiko Nakayama

東北大学グローバルCOE Network Medicine創生拠点冬の合宿2011　2011/02/05
Genome-wide comprehensive and comparative analysis of next-generation sequencing data between ChIP-seq and RNA-seq

Yuichiro Nishida

東北大学グローバルCOE Network Medicine創生拠点冬の合宿2011　2011/02/05
Gemininは造血幹細胞の維持と巨核球の分化を制御する

中野星児, 佐々木陽丞, 本橋ほづみ, 中山啓子

第33回日本分子生物学会年会・第83回日本生化学会大会合同大会　2010/12/07
がん遺伝子RASによる広範囲染色体領域の遺伝子サイレンシング機構の解析

舟山亮, 細金正樹, 西田有一郎, 中山啓子

第33回日本分子生物学会年会・第83回日本生化学会大会合同大会　2010/12/07
Ras-mediated gene silencing によるFas遺伝子領域のエピジェネティック制御

細金正樹, 舟山亮, 西田有一郎, 中山啓子

第33回日本分子生物学会年会・第83回日本生化学会大会合同大会　2010/12/07
Gemininは造血幹細胞の維持と巨核球の分化を制御する

中野星児, 佐々木陽丞, 本橋ほづみ, 中山啓子

第33回日本分子生物学会年会・第83回日本生化学会大会合同大会　2010/12/07
Cdt1 inhibitor, Geminin negatively regulates thrombopoiesis

中山啓子

細胞周期ミニシンポジウム　2010/11/09
Geminin negatively regulates thrombopoiesis

Keiko Nakayama

MEXT Priority Research Project “Cell Proliferation Control” International Symposium Cell Cycle and Cell Differentiation From A to Z　2010/11/04
がん遺伝子Rasは複数の遺伝子を含む広範囲な染色体領域を抑制する

細金正樹, 舟山亮, 中山啓子

日本分子生物学会第10回春季シンポジウム　2010/06/07
Ras-mediated regional silencing around Fas gene Locus. International-presentation

Masaki Hosogane, Ryo Funayama, Jafer Sharif, Haruhiko Koseki, Keiko Nakayama

COLD SPRING HARBOR ASIA CONFERENCES, Epigenetics, Chromatin & Transcription　2010/05/18
核内IKKβを介したNFーkB活性化による細胞死亢進

土谷佳弘, 浅野知一郎, 中山啓子, 加藤知久, Michael Karin, 鎌田英明

第３２回日本分子生物学会年会　2009/12/09
ユビキチンリガーゼSCFFbxw7はNoｔｃｈタンパク質分解によりB細胞の分化を制御する

青山慧, 石川善則, 小野山一郎, 中山敬一, 中山啓子

第３２回日本分子生物学会年会　2009/12/09
Ku８０のユビキチン化を介したDNA二重鎖切断修復制御

石田典子, 家村俊一郎, 安井明, 夏目徹, 中山啓子

第３２回日本分子生物学会年会　2009/12/09
核内IKKβを介したNF-kB活性化による細胞死亢進

土谷佳弘, 浅野知一郎, 中山啓子, 加藤友久, Michael Karin, 鎌田英明

第３２回日本分子生物学会年会　2009/12/09
ユビキチンリガーゼSCFFbxw７はNotchタンパク質分解によりB細胞の分化を制御する

青山慧, 石川善則, 小野山一郎, 中山敬一, 中山啓子

第３２回日本分子生物学会年会　2009/12/09
T-cell differentiation and proliferation in Geminin gene-deleted mouse

中野星児, 山田秀俊, 村井寛子, 佐竹正延, 中山啓子

第３２回日本分子生物学会年会　2009/12/09
Ku８０のユビキチン化を介したDNA二重鎖切断修復制御

石田典子, 家村俊一郎, 安井明, 夏目徹, 中山啓子

第３２回日本分子生物学会年会　2009/12/09
Differential tissue specificity of ubiquitin ligase SCFFbxw7

Keiko Nakayama

東北大学グローバルCOE Network Medicine創生拠点国際シンポジウム　2009/12/07
Defect of T-cell differentiation and proliferation in Geminin genedeleted mouse

Seiji Nakano

東北大学グローバルCOE Network Medicine創生拠点国際シンポジウム　2009/12/07
Ubiquitin ligase SCFFbxw7 regulates differentiation of B lymphocytes through Notch protein degradation

Satoru Aoyama

東北大学グローバルCOE Network Medicine創生拠点国際シンポジウム　2009/12/07
細胞内ストレスに応答した核内IKKβを介した新規ＮＦ-kB活性化機構

土谷佳弘, 浅野知一郎, 中山啓子, 加藤智久, karin Michael, 鎌田英明

第８２回日本生化学会大会　2009/10/21
Cell cycle control during differentiation: regulation by two F-box protein, Fbw7 International-presentation

Keiko Nakayama

CSI Singapore, NUS-Tohoku University GCOE Joint Symposium　2009/09/09
Differential specificity of substrate accumulation in ubiquitin ligase SCFFBXW7 deficient cells International-presentation

Keiko Nakayama

Institite of Molecular and Cell Biology (IMCB) Cell Cycle Regulation and Tumorigenesis Symposium　2009/09/07
Gemininはmouse Embryonic Stem CellにおけるDNA複製と遺伝子発現を制御する

山田秀俊

第６回東北大学バイオサイエンスシンポジウム　2009/06/16
ユビキチンリガーゼSCFFbxw7はBリンパ球の分化・生存の制御因子である

青山慧

第６回東北大学バイオサイエンスシンポジウム　2009/06/16
タンパク質の翻訳後修飾制御機構とその破綻による疾患発症メカニズムの解明

中山啓子

東北大学Network Medicine創生拠点冬の合宿　2009/02/14
DNA損傷による中心体増幅を起こす経路の特定

山田真生, 田中正和, 中山啓子, 中山敬一, 藤澤順一, 三輪正直

第31回日本分子生物学会年会第81回日本生化学会大会合同大会　2008/12/09
ユビキチンリガーゼＳＣＦＦｂｘｗ７は表皮角化細胞において増殖と分化を抑制する

石川善則, 奥山隆平, 小野山一郎, 青山慧, 中山敬一, 中山啓子

第31回日本分子生物学会年会第81回日本生化学会大会合同大会　2008/12/09
ユビキチンリガーゼＳＣＦＦｂｘｗ７はＢリンパ球の分化・生存の抑制因子である

青山慧, 石川善則, 小野山一郎, 中山敬一, 中山啓子

第31回日本分子生物学会年会第81回日本生化学会大会合同大会　2008/12/09
ノックインマウスを用いたｐ２７とｐ５７の機能的類似性と特異性の検討

洲崎悦生, 中山啓子, 中山敬一

第31回日本分子生物学会年会第81回日本生化学会大会合同大会　2008/12/09
新規ＲＩＮＧ-Ｆｉｎｇｅｒタンパク質はＮＡＰ１Ｌ１のユビキチン化を介して細胞増殖を抑制する

石田典子, 家村俊一郎, 夏目徹, 中山敬一, 中山啓子

第31回日本分子生物学会年会第81回日本生化学会大会合同大会　2008/12/09
新規ＲＩＮＧ‐Ｆｉｎgerタンパク質はＮＡＰ１Ｌ１のユビキチン化を介して細胞増殖を制御する

石田典子, 家村俊一郎, 夏目徹, 中山敬一, 中山啓子

第31回日本分子生物学会年会第81回日本生化学会大会合同大会　2008/12/09
ES細胞におけるGemininの発現消失はPolyploidとTrophectoderm分化を誘導する

山田秀俊, 中山啓子

第31回日本分子生物学会年会第81回日本生化学会大会合同大会　2008/12/09
ユビキチンリガーゼＳＣＦＦbxw7はBリンパ球の分化・生存の抑制因子である

青山慧, 石川善則, 中山啓子

東北大学大学院医学系研究科医科学専攻ルネサンス計画事業第2回リトリート大学院発表会　2008/12/06
ユビキチンリガーゼＦｂｗ１１の発生初期におけるタンパク質分解

中山啓子

平成２０年度特定領域研究「タンパク質分解」班会議　2008/11/26
細胞増殖・分化・死

中山啓子

第６０回日本細胞生物学会大会　2008/06/29
G0-G１期を制御するユビキチンリガーゼと発がん

中山啓子

第72回日本生化学会中央支部例会・シンポジウム　2008/05/24
Differential specificity of substrate accumulation in ubiquitin ligase SCFFBXW7 deficient fibroblasts and keratinocytes International-presentation

Ishikawa,Y, Onoyama,I, Okuyama, R, Nakayama, KI, Nakayama, K

Cold Spring Harbor Laboratory 2008 Meeting & Courses, The Cell Cycle　2008/05/14
Conditional inactivation of Fbxw7 results in a defect in cell cycle exit and tumorigenesis International-presentation

Onoyama,I, Tsunematsu, R, Matsumoto,A, Nakayama,K, Nakayama, KI

Cold Spring Harbor Laboratory 2008 Meeting & Courses, The Cell Cycle　2008/05/14
A p27-p57 knock-in mouse model uncovers common and specific roles of p27 and p57 International-presentation

Susaki,E, Nakayama, K, Nakayama,KI

Cold Spring Harbor Laboratory 2008 Meeting & Courses, The Cell Cycle　2008/05/13
Cell cycle control during differentiation:regulation by two F-box proteins, Skp2 and Fbw7

Keiko Nakayama

The 3rd International Workshop on Cell Regulations in Division and Arrest Under Stress　2008/04/06
ユビキチンリガーゼFbw7の組織特異的な生理的基質および機能の解析

石川善則, 小野山一郎, 奥山隆平, 中山敬一, 中山啓子

東北大学大学院医学系研究科医科学専攻ルネサンス計画事業第１回リトリート大学院発表会　2008/02/16
F-boxタンパク質β-TrCP1によるPeriodタンパク質の制御,

益田邦洋, 石川善則, 柳原徳子, 原賢太郎, 舟山亮, 石田典子, 海野倫明, 中山啓子

東北大学大学院医学系研究科医科学専攻ルネサンス計画事業第１回リトリート大学院発表会,　2008/02/16
Gdx, an X-Linked Ubiquitin-Like Modifier, Is Conjugated to Cyclin F and Regulates Mitosis, International-presentation

N. Ishida, S. Hatakeyama, K. Nakayama, K.-i. Nakayama

4th International Conference, Ubiquitin, Ubiquitin-Like Proteins, and Cancer　2008/02/07
Gemininは着床前初期発生に必須である

原賢太郎, 中山敬一, 中山啓子

CREST第4回公開シンポジウム　2008/01/28
腎障害進行におけるSkp2発現亢進の解析：Skp2KOマウスを用いた検討

鈴木小由理, 深澤洋敬, 北川恭子, 内田千晴, 服部隆行, 磯部智康, 小田敏明, 三崎太郎, 中山啓子, 中山敬一, 菱田明, 山本龍夫, 北川雅敏

第30回日本分子生物学会年会・第80回日本生化学会合同大会　2007/12/11
新規ユビキチン様修飾分子GdXによるcyclin Fと細胞周期M期の制御

石田典子, 畠山鎮次, 中山啓子, 中山敬一

第30回日本分子生物学会年会・第80回日本生化学会合同大会　2007/12/11
ユビキチンリガーゼFbw7は表皮角化細胞において増殖と分化を制御する

石川善則, 奥山隆平, 小野山一郎, 中山敬一, 中山啓子

第30回日本分子生物学会年会・第80回日本生化学会合同大会　2007/12/11
Fbxw7によるG0期維持機構とその破綻による発がん

小野山一郎, 松本有樹修, 中山啓子, 中山敬一

第30回日本分子生物学会年会・第80回日本生化学会合同大会　2007/12/11
Gemininは着床前初期発生に必須である

原賢太郎, 中山敬一, 中山啓子

第30回日本分子生物学会年会・第80回日本生化学会合同大会　2007/12/11
ユビキチンリガーゼFbw7はマウス胎仔線維芽細胞においてNotch依存的な細胞増殖障害を制御する

石川善則, 小野山一郎, 中山敬一, 中山啓子

CREST第4回公開シンポジウム　2007/11/20
Fbw7によるG0期維持とその破綻による発がん

小野山一郎, 松本有樹修, 中山啓子, 中山敬一

CREST第4回公開シンポジウム　2007/11/20
細胞増殖から細胞分化への分岐点を解明する

中山啓子

特定領域研究「細胞周期フロンティア-増殖と分化相関　2007/10/29
発がんとG1-G0期制御に寄与するユビキチンリガーゼ（Abnormalities in Cell Cycle Regulation Oncogenesis and ubiquitin ligase related to G1-G0 transition)

中山啓子, 中山敬一

日本癌学会総会　2007/10/03
Ubiquitin ligases required for the regulation of G0-G1 transision International-presentation

Keiko Nakayama

Office of Life Sciences/National University of Singapore and Tohoku University/Center of Excellence Joint Syposium　2007/09/05
Geminin is essential for the development of preimplantation mouse embryos International-presentation

K. Hara, K.I. Nakayama, K. Nakayama

Office of Life Sciences/National University of Singapore and Tohoku University/Center of Excellence Joint Syposium　2007/09/05
Ｇ0-Ｇ1期を制御するユビキチンリガーゼと発がん

中山啓子

千里ライフサイエンスセミナー　2007/07/04
GdX, an X-linked ubiquitin-like modifier, is conjugated to cyclin F and regulates cell cycle International-presentation

Noriko Ishida, Shigetsugu Hatakeyama, Keiko Nakayama, Keiichi Nakayama

The 3rd International Symposium on "Novel perspectives in cancer research and translation to the clinic"　2006/11/09
Accumulation of Notch induces cell cycle arrest in the tumor suppressor Fbxw7-deficient MEFs International-presentation

Yoshinori Ishikawa, Ichiro Onoyama, Kei-ichi Nakayama, Keiko Nakayama

The 3rd International Symposium on "Novel perspectives in cancer research and translation to the clinic"　2006/11/09
Geminin is essential for the development of preimplantation mouse embryos International-presentation

Kentaro Hara, Keiichi Nakayama, Keiko Nakayama

The 3rd International Symposium on "Novel perspectives in cancer research and translation to the clinic"　2006/11/09
FBW7 is a key regulator of Cell Cycle arrest and Tumorigenesis International-presentation

Keiko Nakayama

The 3rd International Symposium on Novel perspectives in cancer research and translational to the clinic　2006/11/09
ヒト滑膜肉腫関連癌原遺伝子SYTは初期発生に必須である

木村太一, 澤洋文, 中山啓子, 中山敬一, 長嶋和郎, 田中伸哉

第65回日本癌学会学術総会　2006/09/28
p27の増加はDNA損傷後の中心体数異常の抑制に必要である

杉原英志, 豊島秀男, 中山啓子, 中山敬一, 佐谷秀行, 三輪正直

第65回日本癌学会学術総会　2006/09/28
分化における細胞周期調節：その破綻による発がん

中山敬一, 中山啓子

第65回日本癌学会学術総会　2006/09/28
がんと細胞周期分化における細胞周期調節その破綻による発がん

中山敬一, 中山啓子

日本癌学会総会　2006/09/28
細胞周期におけるユビキチン化の役割

中山啓子

第10回Molecular Cardiovascular Conference　2006/09/08
肥満・糖尿病発症におけるユビキチンリガーゼSkp2の役割

坂上浩, 神戸大学大学院医学系研究科応用分子医学講座糖尿病代謝, 消化器, 腎臓病学分野, 酒井太門, 中村武寛, 岡田潮, 岡田裕子, 高島康弘, 永礼智基, 中村

日本臨床分子医学会学術総会　2006/07/20
ユビキチンリガーゼFbw7はMEFにおいて増殖促進因子として

石川善則, 小野山一郎, 中山敬一, 中山啓子

第1回COE相互交流、若手研究者の会　2006/07/14
GEMININ IS ESSENTIAL FOR DEVELOPMENT OF

Kentaro Hara, Keiichi Nakayama, Keiko Nakayama

第1回COE相互交流、若手研究者の会　2006/07/14
Regulation of CDK inhibitors by ubiquitin ligase Fbxw7 International-presentation

Yoshinori Ishikawa, Ichiro Onoyama, Kei-ichi Nakayama, Keiko Nakayama

29th Annual Meeting of the Molecular Biology Society of Japan　2006/06/18
Geminin is essential for development of preimplantation embryos International-presentation

Kentaro Hara, Kei-ichi Nakayama, Keiko Nakayama

29th Annual Meeting of the Molecular Biology Society of Japan　2006/06/18
Gdx, AN X-LINKED UBIQUITIN-LIKE MODIFIER, IS CONJUGATED TO International-presentation

Noriko Ishida, Shigetsugu Hatakeyama, Shun-ichiro Iemura, Toru Natsume, Keiko Nakayama, Keiichi Nakayama

29th Annual Meeting of the Molecular Biology Society of Japan　2006/06/18
GEMININ IS ESSENTIAL FOR DEVELOPMENT OF PREIMPLANTATION EMBROYS International-presentation

Kentaro Hara, Keiko Nakayama, Keiichi Nakayama

Cold Spring Harbor Symposium "The Cell Cycle"　2006/05/17
Geminin is essential for development of preimplantation embroys International-presentation

Keiko Nakayama, Kentaro Hara, Keiichi Nakayama

Cell Regulation in Division and Arrest Workshop　2006/03/06
ｐ27はＤＮＡ損傷後の中心体過剰複製を抑制する

杉原英志, 筑波大学人間総合科学研究科, 斎藤総一郎, 新田孝幸, 豊島秀男, 中山啓子, 中山敬一, 佐谷秀行, 三輪正直

日本癌学会総会　2005/09/14
細胞周期とがん研究の新展開細胞増殖の制御におけるユビキチン化の役割

中山啓子, 中山敬一

日本癌学会総会　2005/09/14
細胞周期とがんＭ期進行におけるｐ27分解の重要性

中山啓子, 三宅智, 中山敬一

日本癌学会　2004/09/29
ＥＩＤファミリータンパクは転写抑制によって分化を阻害する

三宅智, 中山啓子

日本癌学会総会　2004/09/29
31ST INTERNATIONAL SYMPOSIUM ON ALS/MND

An Amyotrophic Lateral Sclerosis-Associated Mutant of, ORF, Is Stabilized, by NEK, Mediated Hyperphosphorylation, the Inability to Bind FBXO

31ST INTERNATIONAL SYMPOSIUM ON ALS/MND　2000/12/11

Show all Show first 5

Research Projects 60

幹細胞における時空間リプログラミングの機構解明と疾患との関わりの研究

中山敬一, 中山啓子

Offer Organization: 日本学術振興会

System: 科学研究費助成事業

Category: 基盤研究(A)

2023/04 - 2026/03
膵癌術前治療耐性克服を目指したcollagen XVIIを標的とする新規治療開発

水間正道, 中山啓子, 舟山亮, 青木修一

Offer Organization: 日本学術振興会

System: 科学研究費助成事業

Category: 基盤研究(C)

Institution: 東北大学

2022/04/01 - 2025/03/31
SETD5の膵臓癌における機能

中山啓子, 舟山亮, 中川直, 細金正樹

Offer Organization: 日本学術振興会

System: 科学研究費助成事業

Category: 基盤研究(B)

Institution: 東北大学

2021/04/01 - 2024/03/31

More details Close

膵臓がん由来のRAS活性化型変異細胞（以降 PancCAと記載）のMEK阻害薬処理によって得られる阻害薬耐性細胞では、SETD5のタンパク質発現量が著しく増加することが報告されていた。まず、この報告の再現性と汎用性の確認を行った。マウス膵臓がん由来のRAS活性化型変異細胞（KPC・AK4.4）を用いて、MEK阻害薬またはHDAC3阻害薬を添加し3日から12日間培養した。形態の変化や増殖の低下が観察されたがSETD5タンパク質蓄積を誘導することができなかった。これまでの報告とは、実験条件が多少異なっているが、一般的にRASの変異により増殖が活性化している腫瘍においてMEK阻害薬抵抗性獲得に、SETD5の発現量の変化は貢献していないと結論付けた。これまでの我々は、神経細胞で、SETD5は細胞増殖に関わる分子の翻訳を制御していることを見出している。そこでRAS活性化型変異細胞においてもSETD5が同様の機能を有するのか調べることとした。CRISPR-Cas9システムを用いて、SDTD5を持たないRAS活性化型変異細胞（AK4.4-SETD5del）を作製した。In vitro の培養系では、AK4.4-SETD5del細胞は、野生型AK4.4に対して明らかな増殖の違いや、MEK阻害薬に対する感受性の違いを見出すことはできなかった。そこで、これらの細胞を同系統のマウス皮下に接種し、in vivo での腫瘍増殖能を調べたところ、AK4.4-SETD5delより発生した腫瘍は、野生型細胞より発生した腫瘍に比して、著しく腫瘍増大率が低下していることを見出した。
Investigation for mechanisms underlying cell cycle regulation and metabolism in stem cells

Offer Organization: Japan Society for the Promotion of Science

System: Grants-in-Aid for Scientific Research

Category: Grant-in-Aid for Specially Promoted Research

Institution: Kyushu University

2018/04/23 - 2023/03/31
Identification of novel microexon involved in the regulation of colorectal cancer

Hideaki Karasawa

Offer Organization: Japan Society for the Promotion of Science

System: Grants-in-Aid for Scientific Research

Category: Grant-in-Aid for Scientific Research (C)

Institution: Tohoku University

2018/04/01 - 2021/03/31

More details Close

Microexons are very small exons, whose alternative splicing is frequently deregulated in a neurological disease, autism spectrum disorder. However, little is known about the splicing pattern, regulatory mechanisms, and roles of microexon in cancer. We have examined the transcriptome-wide profile of microexon splicing in matched normal colon and colorectal cancer (CRC) tissue specimens. Out of 1,492 microexons with shorter than 15 nucleotides in length, seventeen (1%) were differentially spliced between normal colon and CRC tissues. Analyses of RNA sequence motifs, an RNA-binding protein database, and subsequent shRNA-mediated knockdown identified two splicing factors, RBFOX2 and PTBP1, as regulators of microexon splicing. Our data thus suggest that the altered expression of RBFOX2 and PTBP1 might affect cell adhesion and migration of CRC cells through the regulation of microexon splicing.
細胞老化へ向かう代謝アダプテーションのエピゲノムからの探索

中山啓子

System: 日本学術振興会科学研究費助成事業新学術領域研究

2018/04 - 2020/03
過剰な遺伝子増幅を誘導するシグナル経路を明らかにする

中山啓子

System: 日本学術振興会科学研究費助成事業挑戦的研究(萌芽)

2018/04 - 2020/03
ヒストンコードによる転写伸長速度制御機構の解明

中山啓子

System: 日本学術振興会科学研究費助成事業基盤研究(B)

2017/04 - 2020/03
PADI2 suppresses proliferation of colon cancer cells through protein citrullination

Funayama Ryo, Nakayama keiko, TANIGUCHI hajime

Offer Organization: Japan Society for the Promotion of Science

System: Grants-in-Aid for Scientific Research

Category: Grant-in-Aid for Scientific Research (C)

Institution: Tohoku University

2016/04/01 - 2019/03/31

More details Close

PADI2 was expressed in normal colon epithelial cells and downregulated during colon carcinogenesis. Overexpression of PADI2 suppressed proliferation of colon cancer cells in vitro concomitant with increased protein citrullination. The growth defect was not caused by increased apoptosis but by suppression of cell cycle progression. Some citrullinated proteins were found in normal colon tissues. Our data thus suggest that PADI2 suppresses proliferation of colon epithelial cells through protein citrullination, and that downregulation of PADI2 expression might contribute to colon carcinogenesis.
Integrative study for functional analyses of stem cell maintenance factors and visualization of stem cells

Nakayama Keiichi, GOTOH Yukiko

Offer Organization: Japan Society for the Promotion of Science

System: Grants-in-Aid for Scientific Research

Category: Grant-in-Aid for Scientific Research (S)

Institution: Kyushu University

2013/05/31 - 2018/03/31

More details Close

On the basis of our previous studies, we found that 1) low proliferation, 2) low metabolism, 3) low oxidation, which are attributable to p57, Fbw7, and Fbxl5, respectively, are hallmarks of tissue stem cells. The aim of this study is to elucidate the biological significance of p57, Fbw7, and Fbxl5 for stem cell function and maintenance. During the course of this study, we noticed that p57 is the most restricted marker for stem cells. We thus traced the lineage of p57-positive cells in mice, and found that p57 shows a stem cell-specific expression pattern in hematopoietic, neural, and intestinal stem cells. Furthermore, ablation of p57 gene in stem cells impaired the function and maintenance of stem cells with the cell cycle being aberrantly activated. These results thus suggested that cell cycle arrest induced by p57 CDK inhibitor is essential for stem cell function and maintenance. We also demonstrated that p57 is also required for cancer stem cells.
Genome wide analysis of epigenetic regulation of enhancer activity via H3K27 modifications

HOSOGANE Masaki, NAKAYAMA Keiko, FUNAYAMA Ryo, SHIROTA Matuyuki

Offer Organization: Japan Society for the Promotion of Science

System: Grants-in-Aid for Scientific Research Grant-in-Aid for Young Scientists (B)

Category: Grant-in-Aid for Young Scientists (B)

Institution: Tohoku University

2015/04/01 - 2017/03/31

More details Close

It is still unknown whether H3K27me3 histone modification has a repressive role toward enhancer activity. To address this issue, we performed series of genome wide analysis to map and quantify H3K27me3, H3K27ac, p300 and mRNA expression in H3K27me3-depleted K562 human leukemia cells. We observed that the decrease in H3K27me3 level in enhancer region led to massive accumulation of H3K27ac and p300 at the same enhancer region, whereas the decrease in H3K27me3 in gene body was associated with upregulation of transcription of the same gene. Our results thus provide insights into the causal relationship between H3K27 histone modifications in regulatory elements and gene expression.
DNA複製による転写とエピゲノム状態の制御の可能性について

System: 日本学術振興会科学研究費助成事業挑戦的萌芽研究

2016/04 - 2017/03
ヒストン修飾パターンによる転写制御の解明

中山啓子

System: 日本学術振興会科学研究費助成事業基盤研究(B)

2014/04 - 2017/03
Molecular mechanism of oncogenic RAS-induced gene silencing

Funayama Ryo, NAKAYAMA Keiko, NAGASHIMA Takeshi, HOSOGANE Masaki

Offer Organization: Japan Society for the Promotion of Science

System: Grants-in-Aid for Scientific Research Grant-in-Aid for Young Scientists (B)

Category: Grant-in-Aid for Young Scientists (B)

Institution: Tohoku University

2014/04/01 - 2016/03/31

More details Close

Cancer is caused by DNA mutations. Some mutations cause alterations in gene expression, which result in abnormal growth and viability in cancer cells. We uncovered the molecular mechanism of transcriptional repression induced by an oncogenic RAS mutation, one of the most recurrent mutation in cancer. Phosphorylation activity of protein kinase Erk2 is required for RAS-induced transcriptional repression. Moreover, transcriptional repression triggers alteration of chromatin environment including histone modification.
腫瘍形成に求められるゲノム多様性の検証

System: 日本学術振興会科学研究費助成事業挑戦的萌芽研究

2015 - 2016/03
Functional analysis and clinical application of the Ubiquitin - Proteasome pathway in the process of renal ischemia reperfusion injury.

MOTOYOSHI NAOTAKA, SAIKI Yoshikatsu, KAWAMOTO Shunsuke, ABE Takaaki, NAKAYAMA Keiko, OKAMURA Masashi

Offer Organization: Japan Society for the Promotion of Science

System: Grants-in-Aid for Scientific Research

Category: Grant-in-Aid for Scientific Research (C)

Institution: Tohoku University

2012/04/01 - 2015/03/31

More details Close

We have established the porcine renal ischemia-reperfusion injury model clamping bilateral renal arteries. The pigs were allocated in the two groups; control and Bortezomib (proteasome inhibitor) administration group. We assessed the degrees of kidney damage and expression of PPAR-γ and LXR-α (PPAR-γ’s target gene) in the kidney comparing the Bortezomib administration group with control group. Serum Neutrophil gelatinase-associated lipocalin (NGAL) was used as an indicator of acute kidney injury. Histopathological changings were evaluated by the pathological scoring. Qualification and quantification of the expression of PPAR-γ, LXR-α, and downstream regulatory factors(NFκB, AP-1, NFAT and STAT-3) were evaluated by immunohistochemical staining and real-time RT-PCR method.
ヒストン修飾酵素の機能に関する網羅的スクリーニング

中山啓子

System: 日本学術振興会科学研究費助成事業挑戦的萌芽研究

2014/04 - 2015/03
Analysis of carcinogenic mechanism of biliary tract cancer from the protein control by ubiquitin ligase Fbw7

KATAYOSE Yu, YAMAMOTO Kuniharu, YOSHIDA Hiroshi, NAKAGAWA Kei, HAYASHI Hiroshi

Offer Organization: Japan Society for the Promotion of Science

System: Grants-in-Aid for Scientific Research

Category: Grant-in-Aid for Scientific Research (B)

Institution: Tohoku University

2010 - 2012

More details Close

The gene variation of Fbw7 of a biliary tract cancer cell line was considered, three sorts of isoform(s) of alpha-beta-gamma existed, and especially the expression level of gamma traced that it was low in predominance by all the cell strains in order to solve the role of Fbw7 in a biliary tract cancer occurrence. Next, in order to solve protein control of a biliary tract cancer, the Mouse Embryonic Fibroblast (MEF) of Fbxw7 conditional KO (CKO) mouse, CDK inhibitor showed clearly to have received the complicated control by the proteolytic mechanism by Fbxw7.
RAS活性化による転写制御ー染色体領域に注目したアプローチによる解析

中山啓子

Offer Organization: 日本学術振興会

System: 科学研究費助成事業

Category: 基盤研究(B)

Institution: 東北大学

2011 - 2011
In vivoタンパク質複合体形成モニタリングの試み

中山啓子, 石田典子, 舟山亮, 中野星児

Offer Organization: 日本学術振興会

System: 科学研究費助成事業

Category: 挑戦的萌芽研究

Institution: 東北大学

2010 - 2011

More details Close

マウス個体(特に発生過程)でのタンパク質の相互作用変化について高い時空間的分解能を持った解析法を確立することを目的に本研究を行った。タンパク質はそれが相互作用する分子により、異なる機能発揮するが、相互作用分子の研究は試験管内で細胞抽出液中を用いた試験管内での結合を調べるに過ぎなかった。そこで、本研究課題では、in vitroで相互作用することが知られている分子群が、in vivoでは、いつ、どこで相互作用するのかを半定量的にモニターする方法を開発することを目指し、発生工学的手法を用いて蛍光タンパク質の断片との融合タンパク質を発現するマウスを作製、in vivoでタンパク質複合体の挙動をモニタリングする。本研究ではモデルタンパク質としてGemininを用いた。 GemininはCDT1とHoxB9に結合することが知られている。そこで、Geminin・CDT1・HoxB9をそれぞれ異なる蛍光タンパク質との融合タンパク質を作製する。これらの融合タンパク質がこれまでに報告されている機能を持つことを培養細胞内で確認するために、NIH3T3細胞にこれらの融合タンパク質を過剰発現し、細胞周期に与える影響と、これまで報告されている結合タンパク質との結合を免疫沈降法で確認した。また、これらの融合タンパク質を共発現し、想定される蛍光がみられることを確認した。以上の結果を踏まえて、in vivoでのモニタリングを行うために、lck promoterによって各融合タンパク質のcDNAが転写されるような発現ベクターの構築を終えた。
The turning point from cell proliferation to cell differentiation

NAKAYAMA Keiko

Offer Organization: Japan Society for the Promotion of Science

System: Grants-in-Aid for Scientific Research

Category: Grant-in-Aid for Scientific Research on Priority Areas

Institution: Tohoku University

2007 - 2011
Analysis of M phase-regulation by GdX

ISHIDA Noriko, NAKAYAMA Keiko, FUNAYAMA Ryo, NAKANO Seiji

Offer Organization: Japan Society for the Promotion of Science

System: Grants-in-Aid for Scientific Research

Category: Grant-in-Aid for Scientific Research (C)

Institution: Tohoku University

2008 - 2010

More details Close

We found that cyclin F was covalently conjugated with GdX (GdXylation). Our data suggested that GdXylation of cyclin F regulated its protein stability and cell cycle progression. Although we identified 11 GdX- and 23 RNF (candidate of GdXligase)-binding proteins by biochemical techniques, identification of E1, E2 and E3 enzymes for GdXylation could not be achieved.It was suggested that RNF is functional as a ubiquitinligase in mouse spermatogenesis by analysis of RNF-deficient mice. So we will develop the functional analysis of RNF in future.
Molecular and temporal-spatial regulation of ubiquitin ligase , F-box protein

NAKAYAMA Keiko, ISHIDA Noriko, FUNAYAMA Ryo, NAKANO Seiji, DEL CARPIO Munoz Carlos Adriel, NISHIDA Yuichiro

Offer Organization: Japan Society for the Promotion of Science

System: Grants-in-Aid for Scientific Research

Category: Grant-in-Aid for Scientific Research (B)

Institution: Tohoku University

2008 - 2010

More details Close

F-box protein functions as a ubiquitin ligase constructing SCF protein complex. In this preject, I focused my interest on typical F-box proteins, Fbxw7 and β-TrCP. I analyzed their gene targeting mice to elucidate the biological function of these proteins. Although Fbxw7 suppressed tumorigenesis in T lymphocytes and hematopoietic stem cell, it facilitated cell proliferation in primary embryonic fibroblasts and keratinocytes. b-TrCP promoted cell proliferation.
細胞周期と初期発生に関わる分子群の新規解析法の開発

中山啓子, 舟山亮, 山田秀俊, 原賢太郎

Offer Organization: 日本学術振興会

System: 科学研究費助成事業

Category: 萌芽研究

Institution: 東北大学

2007 - 2008

More details Close

核膜を標識するためにラミンBを、中心体の標識にセントリン2を、細胞膜の標識にβ-カテニン、E-カドヘリンを用いた積まず、これらのタンパク質のcDNAをRT-PCRにて取得し、そのcDNAの5'側または3'側に蛍光色素をコードするcDNAをin frameで連結し、標識タンパク質と蛍光色素の融合タンパク質をコードするcDNAを作製した。融合タンパク質のcDNAを発現ベクターにクローニングし、腫瘍細胞へ発現し、蛍光顕微鏡にて観察し、融合タンパク質が、予想された目的とする細胞内局在をとることを確認した。このベクターを鋳型にRNA polymeraseを用いて転写反応を行い、RNAを作製した。さらにpoly Aを付加した後、精製し、注入用RNAとした。このRNAをマウス一細胞期胚に注入した。RNA注入後、48時間後に蛍光顕微鏡で観察し、期待される細胞内局在をとっているか、胚の発生は正常に進行しているかを判定した。さらにこの胚を固定し、各小器官に特有のタンパク質に対する抗体で、免疫染色を行い、発現させた融合タンパク質と共局在することを確認した。その結果、ラミンBおよびE-カドヘリンは、有用なツールであることが判明したが、中心体を標識するセントリン2は、発現が弱く明瞭な染色が得られなかった。ラミンBおよびE-カドヘリンは、さらに継時的に観察をタイムラプス蛍光顕微鏡で行った。RNA注入12時間後より、30分ごとに96時間までの観察を行い。この間の蛍光強度の変化を調べたところ、いずれも96時間の間に蛍光強度は減衰するものの、観察は可能であることが判明した。
発生におけるタンパク質分解制御機構とその意義の解明

中山啓子, 石田典子, 舟山亮, 山田秀俊, 原賢太郎

Offer Organization: 日本学術振興会

System: 科学研究費助成事業

Category: 特定領域研究

Institution: 東北大学

2007 - 2008

More details Close

β-TrCP1ノックアウトマウスは、正常に産まれ、生命予後に異常はなく、正常に子孫を維持することが可能であった。β-TrCP2ノックアウトマウスは、胎生8.5日より成長が停止し、胎生9.5日では、心嚢や頭部のバルーン状の拡張が見られた。そして、血管の内皮細胞の免疫染色からリモデリング不全が認められた。そこで、胎生8.5日目のβ-TrCP2ノックアウトマウスを詳細に観察するために、病理切片による免疫染色を行った。その結果、β-TrCP2ノヅクアウトマウスは、細胞周期の異常は認められないものの、特に胚後部に強くアポトーシスが観察された。この部位は、レポーターマウスを用いた際に観察されるβ-cateninの活性が高い部位と一致していた。 β-TrCP1とβ-TrCP2の相補性を調べるためには、ダブルノックアウトマウスの表現型の観察を試みた。その結果、ダブルノックアウトマウスは、胎生7.5日目には、観察されるものの、明らかにβ-TrCP2シングルノックアウトマウスに比較し矮小であった。このことは、β-TrCP1とβ-TrCP2の間に相補性があることを示している。しかしながら、胎生7.5日目の死亡原因については、未だ不明である。また、β-TrCP1とβ-TrCP2の相補性を調べるためには、ダブルノックアウトES細胞の樹立を試み、β-TrCP2のノックアウトES細胞の樹立には成功した。β-TrCP2ノックアウトES細胞は、試験管内分化誘導系によって、神経前駆細胞へ分化できることを確認した。また、ヌードマウスに移植し、奇形腫形成能を調べたところ、三胚葉へ分化できることが観察され、β-TrCP2は、何らかの特異的な胚葉形成に必須な分子では無いことが判断した。
Cell Cycle Regulation of Embryogenesis Competitive

2004/01 - 2007/03
Analysis of G0 phase-specific large complex including p27

ISHIDA Noriko, NAKAYAMA Keiko, HARA Kentaro

Offer Organization: Japan Society for the Promotion of Science

System: Grants-in-Aid for Scientific Research

Category: Grant-in-Aid for Scientific Research (C)

Institution: Tohoku University

2006 - 2007

More details Close

We have tried to identify the components of GO-specific large complex including p27 by analytical method for detection of difference between asynchronous and growth arrested in GO phase culture of HEK293T cells. Previously, we reported that the abundance of p27 was decreased in many organs, including brain, thymus. spleen and testis of p27^<Ser10A/Ser10A> knock-in mice. The stability of p27 in G0 phase was markedly reduced in lymphocytes of p27^<Ser10A/Ser10A> mice compared with wild-type cells. Then, we analyzed p27 in G0 phase by size fractionation using gel-filtration and obtained that p27 formed large complex specifically in G0 phase inhibitor. By affinity column chromatography using anti-FLAG antiboby and gel-filtration chromatography, we recovered a free-FLAG-p27, and could collect FLAG-p27 interacting proteins. Then we succeeded to identify 26 p27 interacting proteins (including 13 novel and 13 already-known) in total. Whereas we tried to identify the proteins were associated to p27 specifically in G0 phase by comparison of proteins between asynchronous and growth arrested culture cells, we could not succeed. However, we identified PP2A subunit A (Protein Phosphatase 2A) as a novel binding protein of p27 plays an important role for the stability of p27 becomes to be unstable. Based on this hypothesis, we are examining the effects of PP2A on stability of p27 using phosphatase inhibitor or siRNA against PP2A. We also plan to identify the G0-specific binding proteins of p27 using NIH 3T3, like as normal cells.
Search for physiological substrates and functional analysis of F-box protein β-TrCP in vivo

NAKAYAMA Keiko, ISHIDA Noriko, HARA Kentaro

Offer Organization: Japan Society for the Promotion of Science

System: Grants-in-Aid for Scientific Research

Category: Grant-in-Aid for Scientific Research (B)

Institution: Tohoku University

2006 - 2007

More details Close

Our purpose of this project is elucidation of the regulation of protein expression on development. We have analyzed the function of F-box proteins β-TrCP2, and demonstrated its role on development and differentiation during embryogenesis. β-TrCP1 is the ubiquitin ligase which ubiquitinates β-catenin and l_<κ>βα The gene-targeted mouse of β-TrCP1, which we have generated, still exhibited degradation of β-catenin and l_<κ>βα, suggesting existence of other ubiquitin ligases for β-catenin and l_<κ>βα. The gene-targeted mouse of β-TrCP2, the homologue of β-TrCP1 is embryonic lethal at embryonic day9.5. This difference of phenotype between β-TrCP1 knockout mouse and that of β-TrCP2, indicate that each proteins have different biological functions significantly, even though the difference of biochemical functions have not been reported. We analyzed four proteins picked up based on the expression during early and middle embryogenesis and the consensus sequences, which are recognized by β-TrCP2. However, we did not detected bindings between these proteins and β-TrCP2, and ubiquitination and degradation by β-TrCP2. We have generated β-TrCP2 conditional knockout mouse. B-TrCP1 knockout primary embryonic fibroblasts have shown the abnormal regulation of Period 2 protein expression, suggesting that maintenance of circadian rhythm requires protein degradation by β-TrCP1.
ユビキチンリガーゼCdc4/Fbw7変異にともなう組織特異的細胞増殖機構の解明

中山啓子, 松本雅記

Offer Organization: 日本学術振興会

System: 科学研究費助成事業

Category: 特定領域研究

Institution: 東北大学

2006 - 2007

More details Close

ユビキチンリガーゼCdc4/Fbw7は、Notch、サイクリンE、c-Mycなど細胞増殖を促進する分子をユビキチン化するリガーゼであることが、試験管内の研究で明らかとなっている。また、様々ながんでCdc4/Fbw7の変異が報告されがん抑制遺伝子であると考えられる。すなわちCdc4/Fbw7の機能不全によりその基質分子が異常に蓄積し、そのために異常な細胞増殖、がん化を引き起こしていると考えられる。そこでわれわれは、Cdc4/Fbw7のコンディショナルノックアウトマウスを作製し、Cdc4/Fbw7の生物学的な機能、特に発がんに対する関与を調べた。まず、胎仔線維芽細胞でCdc4/Fbw7遺伝子を欠失させた。すると予想に反して細胞の増殖が著しく抑制された。これは、細胞周期のG1期での停止とアポトーシスの増加によるものであった。その際に、タンパク質の発現量を調べてみると、Cdc4/Fbw7の基質と報告されているもののうち、Notch-1が異常な蓄積を示すものの、サイクリンEやc-Mycの蓄積は認められなかった。そこで、Notch-1が転写因子として機能する際の共役因子であるRBP-jとのダブルノックアウトを作製しNotch-1の過剰な機能発現をキャンセルすることを試みた。すると、Cdc4/Fbw7シングルノックアウトで観察された増殖抑制は解除された。このことから、Cdc4/Fbw7は胎仔線維芽細胞においては、Nocthのシグナル量を制御することによって適切な増殖能力を維持していること考えられた。
新規分化抑制因子EIDによる哺乳類幹細胞の未分化性維持機構の解明

中山啓子, 石田典子, 原賢太郎

Offer Organization: 日本学術振興会

System: 科学研究費助成事業

Category: 特定領域研究

Institution: 東北大学

2005 - 2006

More details Close

EID-1は分化に関わる転写コアクチベーターp300に結合し、p300のヒストンアセチル化酵素(HAT)活性を阻害する。この阻害活性により、p300依存的に分化を抑制した。分化誘導可能なC2C12細胞にEID-1を安定的に過剰発現し、分化に与える影響を調べた。野生型C2C12は、血清除去などによって分化し細胞形態の変化、分化マーカータンパク質の発現が見られるが、EID-1過剰発現C2C12では、分化が阻害された。さらにEID-1はp300のみならずその基質であるピストンにも結合し、そのアセチル化を阻害することによって細胞分化に必要な転写因子の機能を阻害した。一方、EID-1は細胞増殖を停止し分化誘導が開始されると同時に、急激にプロテアソーム依存的分解によって分解された。この分解によって分化関連転写因子の阻害作用は消失し、分化が開始すると示唆された。一方、EID-1の類縁タンパク質として同定したEID-2およびEID-4は、EID-1と異なり転写抑制に働くピストン脱アセチル化酵素(HDAC)と結合し、機能的に協調することによって、分化に関わる転写を阻害する。すなわち結合蛋白は全く異なる機能を持つもののEID-2およびEID-4もEID-1と同様に分化誘導可能な細胞株に過剰発現させると、血清除去などによるMyoDなどの転写活性誘導が見られず、分化誘導が阻害された。さらにマウス筋芽細胞株C2C12細胞への剰発現では、アポトーシスの誘導がみられ、EIDファミリーは単に分化の決定因子として機能するのではなく、アポトーシスも含む細胞の運命決定因子として機能していると考えられた。
タンパク質の安定性に関わる新規翻訳後修飾、Gdx化システムの分子機構の解明

石田典子, 中山啓子

Offer Organization: 日本学術振興会

System: 科学研究費助成事業

Category: 特定領域研究

Institution: 東北大学

2005 - 2006

More details Close

ユビキチンやユビキチン様タンパク質(UBLs)による翻訳後修飾は、生命現象に必須なタンパク質制御機構である。われわれはこれまでにII型 UBLグループに分類されていたGdX/UBL4が新規翻訳後修飾分子(1型 UBL)として機能することを示唆するデータを得ていた。GdXの結合タンパク質として同定したcyclin Fが細胞内でGdX化することを示すデータを得たのである。GdX化を受けないcyclin F変異体を細胞に発現させたり、siRNAによってGdXタンパク質の発現量を減少させるとcyclin Fは安定化し、どちらの場合もG2/M期の細胞が増加した。そこで本特定領域研究期間に、より詳細な細胞周期の解析と新規GdX化システムの酵素群(E1(GdX活性化酵素)/E2(GdX結合酵素)/E3(GdXリガーゼ)様分子)の同定を試みた。まず細胞周期解析のために核を可視化したHeLa細胞を用い、shRNAによる誘導型GdX発現抑制細胞株を樹立し、M期の詳細な解析を行った。GdX発現抑制細胞では細胞増殖が低下し、M期の障害(中期形成異常による染色体分配異常や紡錘体形成異常)による多核細胞や分裂期細胞死が増加するという結果を得た。正常な細胞ではS期からM期にcyclin Fの発現量は高くなるが、GdX発現抑制細胞ではそのcyclin Fの分解が遅延し、その結果cyclin Aやcyclin Bの分解時期が遅れていた。Cyclin Bの分解異常により細胞質分裂が完了せず、細胞が2核になることが報告されていることから、GdXはcyclin Fを修飾(GdX化)することによってcyclin Fタンパク質の安定性を制御し、cyclin Aやcyclin Bを制御することが示唆された。このことよりGdXによるcyclin Fの制御が正常な細胞周期の維持に関わっている可能性が強く示唆された。 GdX化システムに関しては、E1(GdX活性化酵素)/E2(GdX結合酵素)様分子の同定には至らなかった。しかしcyclin FのGdX化には直接関与しないGdX化のE3(GdXリガーゼ)様分子や新たな基質の候補遺伝子を複数同定することに成功した。それらの解析を含め、細胞周期M期の制御に重要なGdX化システムの分子メカニズム、特にE1/E2様分子の同定が今後の課題である。
発がんにおけるユビキチンリガーゼCdc4/Fbw7基質分子の役割

中山啓子, 恒松良祐

Offer Organization: 日本学術振興会

System: 科学研究費助成事業

Category: 特定領域研究

Institution: 東北大学

2005 - 2005

More details Close

F-boxタンパク質Cdc4は、Skp1、Cul-1、Rbx1とともにSCF複合体を構成し、ユビキチンリガーゼとして機能すると考えられている。われわれが作製したCdc4ノックアウトマウスは、Notch-4の異常蓄積による血管形成異常が起こり、胎生10.5日目で死亡した。このことは、胎生期における生理的かつ冗長性を示さない基質分子はNotch-4であると結論された。 Tリンパ球特異的にCdc4を欠損するコンディショナルノックアウトマウスを作製し、T細胞分化過程をCD4およびCD8の発現パターンから解析したところ、分化の中間段階であり、CD4/CD8いずれもが発現し、正の選択・負の選択が行われるステージの細胞が野生型に比べ有意に増加していることが判明した。このステージでは通常増殖を停止しるが、Cdc4ノックアウトT細胞では激しい増殖がこのステージでも認められ、過剰増殖がこの細胞群の増加の原因であると考えられた。また、生後8週以降よりリンパ腫の発症が認められ、Cdc4は当初報告されたとおり、がん抑制遺伝子として機能していることが裏付けられた。このような過剰増殖しているリンパ球の生化学的解析では、Notchファミリーとc-Mycの過剰な蓄積が観察された。そこでNotchが転写因子として働く際の共同因子であるRBP-JκノックアウトマウスとCdc4ノックアウトマウスを交配した。このマウスはNotchシグナルが欠失し、かつc-Mycが蓄積している環境を作製することとなり、それらの分子のTリンパ球増殖の効果を調べたところ、このようなTリンパ球でも異常増殖が見られた。このことからCdc4ノックアウトTリンパ球はc-Mycの過剰蓄積により異常な増殖をきたし癌化が誘導されていると結論した。
分化と増殖の転換の分子機構の解明

三宅智, 中山啓子, 石田典子

Offer Organization: 日本学術振興会

System: 科学研究費助成事業

Category: 基盤研究(C)

Institution: 東北大学

2004 - 2005

More details Close

本年度の研究の結果、以下の成果を得た。 1 EID-2類縁分子として新たにEID-2-1ike inhibitor of differentiation-3(EID-3)をクローニングした。これによってEID-1,-2,-3からなる新しい分子ファミリーという概念を提唱することができた。 2 EID-3はEID-2と同様に、過剰発現系において細胞分化を阻害することを明らかにした。特にマウス筋芽細胞株C2C12を用いた系では、発現レベルの高いものでは細胞死を来すこともわかった。 3 EID-3はEID-2と同様に、ヒストン脱アセチル化酵素(HDAC)と結合し、分化に関わる転写因子(MyoDなど)の転写活性を抑制することを明らかにした。HDACファミリーの中でも特にクラス1HDAC(HDAC1,2)と結合することもわかった。 4 EID-3は核内に局在し、この機能はHDACとの結合および転写阻害機能と共通のドメインを利用していることを明らかにした。 5 EID-2およびEID-3がEID-1と同様に、癌抑制遺伝子産物pRBと細胞内で複合体を形成するすることを証明した。 6 EID-2およびEID-3遺伝子は同一染色体上の近傍に位置し、この両方を含むアンプリコンがある種の癌細胞株で増幅していることがわかった。以上をまとめると、1から5の結果によって、EID-2およびEID-3の生化学的・生物学的機能をよりいっそう明らかにすることができた。また5,6の結果から、EID-2およびEID-3の発がんにおける役割が推測された。
Functional Analysis of Proteins regulated by Ubiquitin Ligase

NAKAYAMA Keiko, ISHIDA Noriko, HARA Kentaro

Offer Organization: Japan Society for the Promotion of Science

System: Grants-in-Aid for Scientific Research

Category: Grant-in-Aid for Scientific Research (B)

Institution: Tohoku University

2004 - 2005

More details Close

We have tried to identify physiological substrates of ubiquitin ligase by establishment of comprehensive-analytical method for detection of difference between before and after genetic engineering in knock-out mice. First, we purified ubiquitinated protein specifically by anti-ubiquitin antibody. Ubiquitin is an abundant protein and exist as free form (not conjugated or binding form in mammalian cells. By affinity-chromatography using a conventional antibody, we recovered a free-ubiqitin mainly, and could not collect ubiquitin conjugated proteins. For improvement of the efficiency of collection, we assesed many kinds of antibodies and finally decided to use the FK2 monoclonal antibody, which binds only substrate-conjugated ubiqitin specifically. We succeeded in concentrating ubiquitinated proteins by affinity-chromatography using FK2 binding column. For comparison of proteins between knock-out mice and wild type mice, we have to choose the most adequate tissue and the most effective method for cell-extraction. We identified the tissue which expressed F-box protein we were interested in by in-situ hybridization. Based on these basic informations, we prepared Fbw7-deficient-embryonic fibroblasts and also wild-type fibroblast, and extracted whole cell lysate from each samples. Cell lysate loaded on column packed FK2-antibody conjugated beads for concentration of ubiquitinated proteins. We plan to compare the difference of the ubiquitinated proteins between Fbw7-deficient and wild-type embryonic fibroblasts.
Gemininノックアウトマウスを用いた複製開始機構の解析

中山啓子, 原賢太郎

Offer Organization: 日本学術振興会

System: 科学研究費助成事業

Category: 特定領域研究

Institution: 東北大学

2004 - 2005

More details Close

GemininはDNA複製のライセンシング因子であるCdt1と結合し、Cdt1の前複製複合体(preRC)との結合を阻害する分子である。GemininがAPCによって分解されることによってCdt1は遊離しpreRCと結合することによって複製が再開すると考えられている。われわれはgemininノックアウトマウスを作製し胎生3.5日胚ですでに異常な形態をしめしていることを昨年までの研究で示してきた。そこで本年は、胎生3.5日胚の観察を継続すると同時に、コンディショナルノックアウトマウスの作製に着手した。 Gemininノックアウトマウス3.5日胚は、細胞および核の大きさが不同であり、また一つの細胞に複数個の核が観察された。さらにBrdUラベルによるDNA合成期の核を染色すると大小不同に核がいずれも染まり、また一つの細胞内でも複数個の核が複製していることが判明した。一方、抗リン酸化ヒストンH3抗体によるM期細胞の染色では、野生型には複数個の細胞が染まるもののgemininノックアウト胚では染色が見られなかった。さらに受精後1.5日目に2細胞期胚を卵管より採取、培養を行った。するとgemininノックアウト胚は、4細胞期胚までは野生型と同様に増殖を継続するものの8細胞期で増殖を停止し、いったん観察される細胞間接着も低下することが判明した。このとき、抗geminin抗体で免疫染色を行うと、2細胞期胚では核内にgemininは染色されるが、ノックアウト胚では、4細胞期よりその染色性が低下し、8細胞期では認められなくなることが判明した。このことから、おそらくは母性由来のgemininが4細胞期までの細胞増殖を支持しているが、それがこの時期に消失すると同時に、過剰なS期の進行と共にM期の進行が阻害が生じていると考えられる。
サイクリンEとMycの分解に関与するユビキチンリガーゼCdc4の発癌に果たす役割

中山啓子

Offer Organization: 日本学術振興会

System: 科学研究費助成事業

Category: 特定領域研究

Institution: 東北大学

2004 - 2004

More details Close

F-boxタンパク質Cdc4は、Skp1、Cu1-1、Rbx1とともにSCF複合体を構成し、ユビキチンリガーゼとして機能すると考えられている。Cdc4は当初サイクリンEのユビキチンリガーゼとして報告されたが、われわれが作製したノックアウトマウスでは、サイクリンEの蓄積は認められずむしろNotch-4の異常蓄積による血管形成異常が認められた。培養細胞を用いたでは、Cdc4はc-Mycにリン酸化依存的に結合し、Cdc4の過剰発現ではc-Mycタンパクの半減期を減少させ、逆にsiRNAiによるノックダウンでは半減期を増加させた。また、Cdc4のSkp1結合部位であるF-boxに変異を導入したCdc4は、c-Mycの半減期を延長した。このようなことから、Cdc4は、c-Mycのユビキチンリガーゼとして働いていることが判明した。一方、Tリンパ球特異的にCdc4を欠損するコンディショナルノックアウトマウスを作製したところ、T細胞の分化については明らかな異常は認めなかったが、末梢のリンパ組織において、増殖障害があり末梢のTリンパ球は抗原刺激に対して応答性が低下していた。また、コンディショナルノックアウトマウスから調製した胎児線維芽細胞においてin vitroでCdc4遺伝子を不活化すると、著しい増殖障害が見られた。生化学的な結果より、c-Mycの過剰蓄積によって、アポトーシスが誘導されている可能性が示唆される。
Analysis of the PKC-δ Signaling Pathway by Proteomics with Embryonic and Genetic Engeneering

NAKAYAMA Keiichi, HATAKEYAMA Shigetsugu, KAMURA Takumi, NAKAYAMA Keiko

Offer Organization: Japan Society for the Promotion of Science

System: Grants-in-Aid for Scientific Research

Category: Grant-in-Aid for Scientific Research (B)

Institution: KYUSHU UNIVERSITY

2003 - 2004

More details Close

The accumulation of genome sequence data has facilitated the establishment of new approaches to systematic characterization of gene expression profiles at the mRNA level (transcriptome). Such strategies do not, however, necessarily provide direct information about the profile of protein expression (proteome), given that the abundance of a given mRNA does not necessarily correlate with that of the encoded protein. Furthermore, numerous characteristics of proteins, including their subcellular localization, interactions with other molecules, stability, and posttranslational modification, are amenable to study only at the protein level. Recent advances in methods for protein identification based on MS and searches of protein or DNA sequence databases have allowed high-throughput analysis of the proteome. Posttranslational modification, including phosphorylation, regulates the functions of proteins by affecting their interactions with other molecules, their enzymatic activity, or their subcellular localization and is pivotal to the control of many cellular processes. Such modification is difficult to identify by standard proteomics approaches, however, because the modified proteins usually constitute a small proportion of all protein molecules. It is therefore necessary first to concentrate such modified proteins and to prevent contamination by highly abundant proteins. Highly sensitive MS analysis is then able to detect various types of posttranslational modification. Protein kinase C (PKC), which comprises 11 closely related isoforms, has been implicated in a wide variety of signaling mechanisms. Among PKC isotypes, PKC-δ is unique in that its overexpression results in inhibition of cell growth. We showed that mice that lack PKC-δ exhibit enlargement of peripheral lymphoid organs, expansion of the B lymphocyte population, as well as the presence of numerous germinal centers in lymphoid tissues in the absence of stimulation. We tried to develop a new approach designated "Proteomics with Embryonic and Genetic Engineering (PGEM)" to uncover the changes in PKC-δ-null mice. In this study, we established efficient and large-scale methods for phosphorylation and ubiquitylation of proteins. Furthermore, we also appled the SILAC method to quantitative analysis of the phosphoproteome.
プロテアソームへのユビキチン化基質のターゲッテング機構の解析

嘉村巧, 中山敬一, 畠山鎮次, 中山啓子

Offer Organization: 日本学術振興会

System: 科学研究費助成事業

Category: 特定領域研究

Institution: 九州大学

2003 - 2004

More details Close

ユビキチン・プロテアソーム系を介したタンパク質分解は細胞周期・シグナル伝達・脳変性疾患等に中心的な役割を果たしている。タンパク質へのユビキチン化反応には、E1,E2,E3の酵素群が関与し、この分野に関する研究は急速に進んでいるが、その後のステップであるポリユビキチン化タンパク質のプロテアソームへのターゲッテング機構に関してはほとんど明らかになっていない。最近、我々は生化学的手法を用いて、KPC1(C末側にRINGフィンガー配列を持つ)とKPC2(N末側にUBL配列、C末側に2つのUBA配列を持つ)の2量体からなる複合体を、CDKインヒビターp27Kip1に対する新たなE3として細胞抽出液より分離・精製した。近年UBLおよびUBA配列をもつタンパク質の機能解析が盛んに行われているが、我々が同定したKPC2に関する報告はいまだなされていない。そこで本研究ではKPC2のユビキチン・プロテアソーム系に対する影響を生化学的、細胞生物学的方法を用いて解析することを目的とする。試験管内および細胞内でKPC2がプロテアソームおよびユビキチンと結合することを確認している。これらの結合にはそれぞれKPC2のUBL及びUBAドメインが必要であった。また試験管内でのp27のユビキチン化反応に対してKPC2のSTI1ドメインが必要であることを明らかにした。さらにRNA干渉法をもちいて細胞内でp27の分解にKPC2が関与していることを明らかにした。またKPC2のノックアウトマウスの作製し、個体レベルでのKPC2の影響を検討することとした。KPC複合体とプロテアソームとの関係が明らかになれば、ポリユビキチン化タンパク質のプロテアソームへのターゲッテング機構解明の手がかりとなると考えられる。
Study of molecular mechanisms of CD4 silencing and role s of Runx proteins during lymphocyte development

TANIUCHI Ichiro

Offer Organization: Japan Society for the Promotion of Science

System: Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

Category: Grant-in-Aid for Scientific Research (B)

2003 - 2004

More details Close

The aim of this research is to understand the molecular mechanisms of CD4 silencing and function of Runx transcriptional factors during lymphocyte development. Because a main experimental approach was in vivo analyses of gene manipulated mice, we have generated several mutant mouse strains including gene targeted mice and transgenic mice. By using gene targeting in Es cells and Cre-loxP system, we have generated flox mutant allele for Runx1,Runx3 or CBFβ gene for conditional inactivation by induction of Cre recombinase by tissue and stage specific manner. We have also generated Runx3 transgenic mouse in which myc-epitope tagged Runx3 was specifically expressed in all αβT lineage cells. Another mutant strain, the Runx1^<Δ446> mouse, in which VWRPY amino acid sequence of Runx1 protein was deleted, was kindly provided. By analyzing these mutant mice, we have obtained following results. 1.iNKT cells are originated from DP thymocytes and require Runx1 function for their development. 2.Runx1 and Runx3 possess redundant function in CD4 silencing and thymocyte maturation. In the double knock out of Runx1 and Runx3, positive selection and thymocyte maturation were severely impaired. 3.The C-terminal VWRPY sequence of Runx1 is essential for CD4 silencing in both immature DN thymocytes and mature CD8-lineage thymocytes. However, VWRPY independent transcriptional repression operate ob another target genes of Runx1. Moreover, the VWRPY motif is essential for Runx1 function in iNKT cells development. 4.Expression of Runx3 by transgene is not sufficient to initiate CD4 silencing. Runx3 would not a molecule that defines lineage specificity of CD4 silencing. 5.CBFβ plays essential roles for Runx complex function during thymocytes differentiation. 6.Binding of Runx1 to three Runx recognition motifs in Eβ enhancer at TCRβ locus is essential for activation of TCRβ gene. However, mutation on one Runx motif out of three result in partial impairment of TCRβ activation in DN thymocytes, but did not affect maintenance of TCRβ activation in mature T cells
サイクリンEのユビキチンリガーゼCdc4が細胞周期進行に果たす役割

中山啓子

Offer Organization: 日本学術振興会

System: 科学研究費助成事業

Category: 特定領域研究

Institution: 東北大学

2003 - 2003

More details Close

サイクリンEは380番目のスレオニン(スレオニン380)がリン酸化されるとタンパクの安定性が低下することから、リン酸化依存的な分解経路があるととが示唆されてきた。2001年にリン酸化スレオニン380を認識するユビキチンリガーゼとしてF-boxタンパクCdc4が報告された。Cdc4の過剰発現によつてサイクリンEの分解が促進されるというデータであった。また、あるがん細胞ラインではCdc4に変異があり、サイクリンEの過剰な蓄積が観察されている。そこでわれわれはCdc4のノックアウトマウスを作製した。Cdc4ノックアウトマウスは胎生10日付近で致死となり、その際、特に頭蓋内出血が多くの個体で認められた。Cdc4ノックアウトマウスでは、原始血管叢は正常に形成されるが、リモデリングが起こらないために、正常の血管形成が認められなかった。死亡原因は、血管形成異常であると考えられた。一方、Cdc4の基質と報告されたサイクリンEは、、Cdc4ノックアウトマウスにおいて量的に変化は認められず、サイクリンEによって制御されるCDK2活性にも変化はみられなかった。このことより少なくとも胎生中期では、サイクリンEの分解にCdc4は必須ではないと結論づけられる。しかし胎生10日胚を用いた解析は生化学的解析などに限界があること、また、サイクリンEの蓄積にともなう発がん機構の解明には成体マウスが必要なことからコンディショナルノックアウトマウスを作製することをめざして研究を進めている。また、Cdc4はがん遺伝子c-Mycもユビキチン化する可能性を示唆するデータが予備的実験から得られている。今後、Cdc4の生物学的役割とともに発がん機構への関与についても検討したい。
リボザイムを用いた組織特異的ノックアウトマウス作製法の開発

中山啓子

Offer Organization: 日本学術振興会

System: 科学研究費助成事業

Category: 萌芽研究

2002 - 2003

More details Close

リボザイムは小さな触媒作用を持つRNAで、1990年代前半より基礎研究で用いられてきたが、特定の遺伝子配列を認識することができるところから、医療等の社会的有用性が大いに期待されてきた。リボザイムはあるRNA上の特定配列を認識し、その部位を切断することによって、遺伝子の発現レベルを低下させることができるRNAである。この反応は、試験管内では非常に効果的に行われるにも関わらず、細胞内での活性は、標的遺伝子に強く依存し一般的な発現抑制システムとして用いることは難しいとされていた。しかし、リボザイムの発現法の工夫によって、細胞内で任意の遺伝子の発現レベルを調節することが可能となった。そこで本研究では我々が持つ発生工学的技術をリボザイムによる発現調節システムと結びつけ、組織特異的なノックアウトマウスの作製、さらにリボザイムライブラリーを用いた網羅的ノックアウトマウスの作製という新しい方法論の確立を目指していた。すでに我々がノックアウトマウスを作製済みのp27やSkp2について検討を行った。まず、従来の発現ベクター及び、レトロレトロウイルスベクター由来を開発し、その有効性の比較検討を行ったところ、レトロウイルスベクターでは安定な遺伝子の導入効率が得られるものの、タンパク発現の抑制効果が低いことが判明した。これは、リボザイムの転写量に依存していると考えている。また、リボザイムの認識配列とその発現低下効果の比較検討を行ったが、一定の法則性を見出すことができなかった。
マウスGemininの発生工学的手法を用いた機能解析

中山啓子, 畠山鎮次, 嘉村巧

Offer Organization: 日本学術振興会

System: 科学研究費助成事業

Category: 特定領域研究

2002 - 2003

More details Close

Gemininは、mitosisの間にだけに分解される分子を網羅的にスクリーングすることによつて発見された分子である。G1期には全く存在しないが、S期からG2期、M期の開始にかけて蓄積し、有糸分裂の中期(metaphase)から後期(anaphase)への移行期に急速に消失する。このタンパク量の急激な変化はAPC/Cによるユビキチン化によって分解を受けるためである。また、Gemininを過剰発現するとDNA複製が抑制されることから、DNA複製のライセンシングに関与する分子であると考えられてきた。このように細胞周期の進行に強く関わっていると考えられる分子Gemininのノックアウトマウスを作製することによって、そのin vivoでの生物学的役割を検討することが本研究の目的である。 Geminin遺伝子をクローニングし遺伝子構造を明らかにした後、Cdt1結合領域をコードするエクソンをネマイシン耐性遺伝子に置換するターゲティングベクターを作製し、定法に則りノックアウトマウスを作製したところ、胎生早期に致死であることが確認された。現在までの解析では、ノックアウト胚は胎生7.5日には観察されていない。このような胎生初期の死亡からも細胞の正常な分裂に必須の分子であることが予想される。一方、2004年には、Gemininは、Polycomb遺伝子との相互作用によって、Hox遺伝子を制御することが報告された。今後は、分化におけるHox遺伝子との相互作用も検討したいと考えている。
プロテアソームへのユビキチン化基質のターゲッテング機構の解析

嘉村巧, 中山敬一, 中山啓子, 畠山鎮次

Offer Organization: 日本学術振興会

System: 科学研究費助成事業

Category: 特定領域研究

Institution: 九州大学

2002 - 2002

More details Close

ユビキチン・プロテアソーム系を介したタンパク質分解は細胞周期・シグナル伝達・脳変性疾患等に中心的な役割を果たしている。タンパク質へのユビキチン化反応には、E1,E2,E3の酵素群が関与し、この分野に関する研究は急速に進んでいるが、その後のステップであるポリユビキチン化タンパク質のプロテアソームへのターゲッテング機構に関してはほとんど明らかになっていない。最近、我々は生化学的手法を用いて、p140 (C末側にRINGフィンガー配列を持つ)とp50 (N末側にUBL配列、C末側に2つのUBA配列を持つ)の2量体からなる複合体を、CDKインヒビターp27Kip1に対する新たなE3として細胞抽出液より分離・精製した。近年UBLおよびUBA配列をもつタンパク質の機能解析が盛んに行われているが、我々が同定したp50に関する報告はいまだなされていない。そこで本研究ではp50のユビキチン・プロテアソーム系に対する影響を生化学的、細胞生物学的方法を用いて解析することを目的とする。現在までに、試験管内および細胞内でp50がプロテアソームおよびユビキチンと結合することを確認している。また試験管内でのp27のユビキチン化反応に対してp50が抑制的に作用することも確認している。そしてp27の分解に対する影響をp50の過剰発現あるいはp50に対するRNA干渉法をもちいて検討中である。さらにはp50のノックアウトマウスの作製中であり、個体レベルでのp50の影響を検討する予定である。p50/p140複合体とプロテアソームとの関係が明らかになれば、ポリユビキチン化タンパク質のプロテアソームへのターゲッテング機構解明の手がかりとなると考えられる。
2種類のサイクリンE分解機構の細胞周期進行における役割

中山啓子

Offer Organization: 日本学術振興会

System: 科学研究費助成事業

Category: 特定領域研究

Institution: 九州大学

2002 - 2002

More details Close

サイクリンEは380番目のスレオニン(スレオニン380)がリン酸化されると、タンパクの安定性が低下することから、以前より知られていた。そこで、このリン酸化を認識するF-boxタンパクをスクリーニングすることによって、F-boxタンパクFbw7を含むがユビキチンリガーゼとしてSCF^<Fbw7>が報告された。我々も、細胞内でFbw7とサイクリンEとの結合がリン酸化依存的に生じ、Fbw7は、少なくとも過剰発現することによってサイクリンEの存在量を制御していることを確認した。そこでFbw7のin vivoにおける役割を検討するためにノックアウトマウスを作製した。Fbw7ノックアウトマウスは胎生10日付近で致死であり、その際、特に頭蓋内出血が多くの個体で認められた。これを詳細に調べるとFbw7ノックアウトマウスでは原始血管叢は正常に形成されるが、リモデリングが起こらず、正常の血管が形成されないことが判明し、Fbw7の胎生致死にいたる原因は血管形成不全による出血死であると考えられた。Fbw7の線虫における相同遺伝子Sel-10は、Notch1及びNotch4に結合しユビキチン化することが報告されていた。そこでこのマウスでのNotchの発現量を調べたところ、Notch4の発現量が有意に増加しており、Fbw7がNotch4のユビキチン化を通じてその発現量をコントロールしている可能性が高く、Notch4-HRT-1系が過剰となり、その結果血管系の分化に障害をきたしていると考えられた。一方サイクリンEはノックアウトマウスにおいても量的に変化せず、その活性も不変であった。
Establishment of therapeutic method for neurodegenerative diseases by specific protein degradation system for abnormal proteinsEstablishment of therapeutic method for neurodegenerative diseases by specific protein degradation system for abnormal proteins

HATAKEYAMA Shigetsugu, YAMASHITA Yoshinori, NAKAYAMA Keiko, NAKAYAMA Keiichi

Offer Organization: Japan Society for the Promotion of Science

System: Grants-in-Aid for Scientific Research

Category: Grant-in-Aid for Scientific Research (B)

Institution: Kyushu University

2001 - 2002

More details Close

Intracellular protein degradation is involved in the regulation of biologically important phenomenon including cell cycle, transcriptional regulation and signal transduction. Ubiquitylation is mediated by a multienzyme cascade and involves the formation by ubiquitin of thiol esters with at least two, and, in most instances, three, distinct types of enzyme: an E1, an E2, and an E3.Then Polyubiquitylated proteins are recognized and degraded by proteasomes. In this cascade, E3 enzymes are thought to determine the substrate specificity of ubiquitylation and have been classified into two families, the HECT and RING-finger families. The prototype U-box protein, yeast Ufd2, was identified as a ubiquitin-chain assembly factor (E4) that cooperates with a ubiquitin-activating enzyme (E1), a ubiquitin-conjugating enzyme (E2), and a ubiquitin-protein ligase (E3) to catalyze ubiquitin chain formation on artificial substrates. The U-box is a domain of -70 amino acids that is present in proteins from yeast to humans. We have reported that six mammalian U-box proteins have now been shown to mediate polyubiquitiylation in the presence of E1 and E2 and in the absence of E3. These data may suggest that U-box proteins constitute a third family of E3 enzymes. Almost U-box proteins interact with molecular chaperones, indicating that U-box type E3 is likely to be a chaperone-dependent E3. To deal with the accumulation of abnormal proteins, molecular chaperones refold abnormal proteins and U-box type E3 seems lo degrade misfolded proteins. Machado-Joseph disease (MJD) is caused by expansion of a polyglutamine tract in the protein MJD1 and this form of MJD1 proteins accumulates in neuron and cause neuronal dysfunction. We have reported that one of the mammalian U-box protein. UFD2a mediated polyubiquitylation of MJD1. We reported that other U-box protein CHIP also is related to Parkinson's disease. We are on going to establish the therapeutic method for neurodegenerative diseases with U-box type E3.
New cancer therapy by degradation of specific proteins

NAKAYAMA Keiichi, YAMASHITA Yoshinori, NAKAYAMA Keiko, HATAKEYAMA Shigetsugu

Offer Organization: Japan Society for the Promotion of Science

System: Grants-in-Aid for Scientific Research

Category: Grant-in-Aid for Scientific Research (B)

Institution: Kyushu University

2001 - 2002

More details Close

We proposed to induce to degradation of specific protein by the modification of ubiquitin-ligase, Recently, it is reported that expression level of many proteins are regulated by not only transcription but degradation rate. Ubiquitin-porteasome system is one of the most useful system to regulate expression level of protein because of its specificity. On this unique point we try to create new ubiquitin-ligase to recognize and induce to degradate proteins which are toxic for our bodies. In this project, we used oncoprotein, Myc as model protein. Because Myc is known to bind to Max, we made several chimeric proteins, Max/Nedd4, Max/b-TrCP1, Max/CHIP. Nedd-4 is a HECT-type ubiquitin-ligase. b-TrCP1 is a F-box proteins which is a component of SCF-type ubiquitin-ligase. CHIP is a U-box type ubiquitin-ligase. It was hypothesized that Myc would be bound and ubiquitylated by these chmeric proteins. We confirmed that this artificial protein Max/CHIP bound to Myc in vitro system and also immunoprecipitation assay in vivo. The half-life of Myc reduced by overexpression of Max/CHIP protein in mammalian cells by expression vector most efficiently. Plaque assays and colony-formation assay shown that Myc-induced transformation is inhibited by Max/CHIP chimeric protein expression. Now, we observe the effect on whole bode. We implanted tumor cells which overexpressed Max/CHIP chimeric protein in Nude mouse to examine the effect of tumorigenesis.
New clearance system of abnormal protein in Polyglutammine disease

NAKAYAMA Keiichi, NAKAYAMA Keiko, HATAKEYAMA Shigetsugu

Offer Organization: Japan Society for the Promotion of Science

System: Grants-in-Aid for Scientific Research

Category: Grant-in-Aid for Scientific Research (B)

Institution: Kyushu University

2001 - 2002

More details Close

Machado-Joseph disease (MJD)/Spinocerebellar ataxia type 3 (SCA3) is neurodegenerative disease which is caused by polyglutamine expansion in a responsible gene product, MJD1/Ataxin3. MJD1 has now been shown to undergo ubiquitylation and degradation by proteasome-dependent pathway. MJD1 with expanded polygiutamine tract was more resistant to degradation than normal MJD1. We established an in vitro system of ubiquitylation of MJD1, thereby biochemically purified activity to mediate polyubiquitylation of MJD1 from rabbit reticulocyte lysate. An AAA-family ATPase VCP was isolated from the active fraction, and found to binds to MJD1. Furthermore, UFD2a, a mammalian ubiquitin-chain assembly factor (E4), associated with VCP and induced polyubiquitylation of MJD1. UFD2a markedly promoted ubiquitylation and degradation of MJD1 with expanded polyglutamine tract, resulting in the clearance of MJD1 protein. In contrast, dominant-negative mutant UFD2a reduced the degradation rate of MJD1, leading to the formation of intracellular aggregation. In Drosophila model, overexpression of UFD2a significantly suppressed the neurodegeneration induced by expression of MJD1 with expanded polyglutamine tract. These findings suggest that E4 is a ratelimiting factor of degradation of pathologic polyglutamine-containing proteins, and may give a potential tool for gene therapy to control the clinical conditions of MJD
中心体過剰複製と染色体倍数性異常の発がんにおける役割の研究

中山啓子

Offer Organization: 日本学術振興会

System: 科学研究費助成事業

Category: 特定領域研究(C)

Institution: 九州大学

2001 - 2001

More details Close

私達は、サイクリンEやp27^<Kip1>のユビキチン依存性をつかさどるユビキチンリガーゼのコンポーネントSkp2をクローニングし、そのノックアウトマウス作製した。Skp2ノックアウトマウスにおいてはユビキチン化の障害の結果1)サイクリンEとp27^<Kip1>の異常な蓄積、2)染色体多倍体化や中心体過剰複製などの異常が観察されたが、個体としての発癌は認められなかった。さらに次の段階として、Skp2ノックアウトマウスを以前に作製したp27^<Kip1>ノックアウトマウスと交配することによってSkp2/p27ダブルノックアウトマウスを作製し、過剰なサイクリンEの蓄積とp27^<Kip1>の消失という癌細胞にしばしば認められる生化学的な異常を再現したが、この個体もやはり発癌は認めなかった。驚いたことに、このSkp2/p27ダブルノックアウトマウスでは、染色体多倍数体化や中心体過剰複製などSkp2ノックアウトマウスでみられた表現型は消失した。このようにSkp2ノックアウトマウスとSkp2/p27ダブルノックアウトマウスは、細胞周期関連蛋白の生化学的特徴は非常に癌細胞と類似しているものの、個体レベルでの発癌はみられず、癌誘発のためにはさらなる異常が必要であることが判明した。また、Skp2ノックアウトマウスより調製されたリンパ球細胞及び胎仔線維芽細胞では、細胞周期のG0期からG1期ではp27は野生型細胞と同様に分解されることが判明した。これによって、p27^<Kip1>タンパク質の量的な調節はSkp2だけではなく、他の因子によっても調節されていることが判明した。
Functional analysis of ubiquitin ligase by knock-out mise

NAKAYAMA Keiko, HATAKEYAMA Shigetsugu, NAKAYAMA Kei-ichi, KITAGAWA Masatoshi

Offer Organization: Japan Society for the Promotion of Science

System: Grants-in-Aid for Scientific Research

Category: Grant-in-Aid for Scientific Research (B)

Institution: KYUSHU UNIVERSITY

2000 - 2001

More details Close

1) We have reported that F-box protein, FWD1 works as a ubiquitin ligase on the degradation of lκB or β-catenin. FWD1 constitutes SCF complex with Cul-1, Skp1, and Rbx1. In this study, we generated knock-out mice of Cul-1 and Skp1, which are components of SCF complex, and another F-box protein, Skp2. Analysis of these mice shows the biological role of protein degradation by SCF complex system, one category of proteolysis by ubiquitin-proteasome system. 2) Skp2 is a ubiquitin ligase of cyclin E and p27^<Kip1>. In Skp2 knock-out mice, cyclin E and p27^<Kip1> was accumulated abnormally. Cells in the mutant mice contain markedly enlarged nuclei with polyploidy and multiple centrosomes, and show a reduced growth rate and increased apoptosis. 3) We generated and characterized mice lacking both Skp2 and p27^<Kip1>. The Skp2^<-/->p27^<-/-> mice did not exhibit the overreplication phenotype, suggesting hat p27^<Kip1> accumulation is required for its development. 4) Cul-1 and Skp1 knock-out mice are died between on embryonic day 5.5 and 6.5. In these embryos, cyclin E is accumulated. It is postulated that Cul-1 and Skp1 are common components of SCF complex and relate to degradation of many kinds of protein. Early embryonic death of knock-out mice supports that SCF complex participate in many kinds of protein degradation system.
Cyclin E過剰蓄積とp27の欠損を伴うマウスを用いた発がん機構の研究

中山啓子, 小南欽一郎

Offer Organization: 日本学術振興会

System: 科学研究費助成事業

Category: 特定領域研究(C)

Institution: 九州大学

2000 - 2000

More details Close

これまでに様々な発癌モデルマウスが作製されてきたが、その多くがリンパ腫など肉腫を好発し、ヒト悪性腫瘍の大部分を占める癌腫を好発するものは少ない。ヒト癌では、細胞周期の進行に必須なサイクリンEが過剰発現し、細胞周期の進行を負に制御するCDKインヒビターp27の発現が低下している例が多くみられる。そこで私達は、細胞周期関連蛋白の生化学的状態をがヒト癌と類似した環境であるp27を持たずサイクリンEが過剰に発現しているマウスを作出し、ヒト発癌のメカニズムの解明とモデルマウスとしての可能性を検討した。昨年、われわれが報告したSkp2ノックアウトマウスは、サイクリンEとp27のユビキチンリガーゼであるSkp2を欠失するマウスであり、サイクリンEとp27の過剰蓄積が観察された。それにともない染色体の倍数性異常、核の増大、中心体の過剰複製が認められた。このマウスをp27ノックアウトマウスと交配しSKp2/p27ダブルノックアウトマウスを作製した。このマウスはp27遺伝子が破壊されているためにp27の蓄積はなく、サイクリンEのみが過剰蓄積したマウスとなる。このマウスではSkp2単独ノックアウトマウスでみられた、染色体倍数性の異常や中心体の過剰複製はみられず、むしろ正常に戻っており、有意な発癌率の上昇も認められなかった。Skp2ノックアウトマウスより調整された胎児線維芽細胞の細胞抽出液を用いて生化学的特性を調べたところ、Skp2ノックアウト細胞ではCdc2活性が低下していることが判明した。このことよりp27の分解抑制による過剩積によるCdc2活性の抑制が染色体倍数性の異常や中心体の過剰複製の原因になっていると結論された。
Identification of the factors mediating ubiquitination of inclusion body in neurodegenerative diseases

NAKAYAMA Kei-ichi, HATAKEYAMA Shigetsugu, NAKAYAMA Keiko, KITAGAWA Masatoshi, KOMINAMI Kin-ichiro

Offer Organization: Japan Society for the Promotion of Science

System: Grants-in-Aid for Scientific Research

Category: Grant-in-Aid for Scientific Research (B).

Institution: Medical Institute of Bioregulation, Kyushu University

1999 - 2000

More details Close

It has been reported that most of inclusion bodies which are found in patients' neurons of neurodegenerative diseases are ubiquitinated. The aim of this research project is to identify the factors to mediate ubiquitination of the proteins constituting the inclusion body. As a model system, we chose Machado-Joseph disease (MJD), in which polyglutamine stretch of MJD1 protein is abnormally expanded. We found that MJD1 protein undergoes ubiquitination both in vivo and in vitro. Using the in vitro ubiquitination system as an assay for detection of ubiquitinating activity, we purified the factors required for ubiquitination of MJD1 with several column chromatography. Finally, the activity was recovered in the fraction > 1,000 kDa with gel-filtration column chromatography. We finally identified some components of the ubiquitinating enzyme complex by amino acid sequencing, and isolated the cDNA encoding the proteins. Currently we examined the biological significance and involvement of the formation of inclusion body in neurodegenerative diseases.
細胞周期調節分子の分解制御に関する研究-マウスSCF複合体の遺伝子クローニングと解析-

畠山鎮次, 中山啓子, 北川雅敏, 中山敬一

Offer Organization: 日本学術振興会

System: 科学研究費助成事業

Category: 特定領域研究(A)

Institution: 九州大学

1999 - 2000

More details Close

細胞内に存在する分子の分解制御は、多方面にわたる医学生物学領域の蛋白分子の機能に重要な働きを果たしている。今まで、いくつかの蛋白分解機構が報告されているが、特に、酵母におけるユビキチン依存性プロテアソーム介在性蛋白分解に関する研究により、Skp1、Cul-1、F-box蛋白複合体(SCF複合体)と言われるユビキチン化を実行する酵素複合体は、細胞周期をはじめ、さまざまな細胞機能に関与する分子の発現調節(分解調節)を行っていることが明らかになった。この構成成分のなかで、F-box蛋白が基質分子をユビキチン化させるための基質特異性を担っていることが推定されている。我々は、酵母におけるアナロジーを利用して、マウスにおけるSkp1、Cul-1及び複数のF-box蛋白を同定した。特に、我々がFWD1/β-TrCP(1つのF boxと7つのWD40リピートを有する)として同定したF-box蛋白は、免疫学上重要であるNF-κBの抑制分子であるIκBαと、細胞接着もしくは癌の分野で注目されるβ-cateninのユビキチン化、及び分解に関与していることが判明した。IκBαとβ-cateninの発現量にかかわるアミノ酸配列に存在するセリン/スレオニンをリン酸化する上流のシグナル及びキナーゼは異なるが、そのモチーフ自体は非常に似ており、リン酸化が起こった場合、FWD1/β-TrCPは、IκBαとβ-cateninのどちらにも結合し、ユビキチン化を遂行する。つまり、SCF^<FWD1/β-TrCP>は、IκBαとβ-cateninに分解されるべきシグナルが入った場合に、それらの分子の分解を実行する中心的なシステムであることがわかった。この結果は、酵母以外の真核細胞におけるSCF複合体によるユビキチン化介在性分解の初めての報告となる。
サイクリン依存性キナーゼ阻害分子p27の分解因子の同定及び解析

小南欽一郎, 中山啓子

Offer Organization: 日本学術振興会

System: 科学研究費助成事業

Category: 特定領域研究(A)

Institution: 九州大学

1999 - 1999

More details Close

CDK阻害分子p27の分解には、ユビキチン-プロテアソーム系の関与が示されているが、どのような分子がそのユビキチン化の特異性を決定するか明らかにされていない。そこで本研究では、p27/Rum1(酵母におけるP27の機能的ホモログ)の分解に関与する因子を取得する目的で、pop1変異株を含む酵母を用いた遺伝学的方法によりそれらの遺伝子群の同定を試みた。 1.p27の分解因子の取得 (1)p27をpop1+変異株中で高発現すると致死の表現型を示すことを見いだした。 (2)酵母で発現するマウス胸腺由来のcDNAライブラリーを作成した。 (3)p27高発現による致死性を抑圧する遺伝子を(2)のcDNAライブラリーより8クローン取得した。 (4)これらには遺伝子発現の抑圧活性が無いことを確かめた。 (5)これらには、ユビキチン遺伝子、60SリボソームL23遺伝子、さらに6つの未知の遺伝子が含まれていた。 (6)現在、それらのp27に対するin vivoでの直接的影響を解析中である。 2.Rum1分解に関わる変異株および阻害因子の取得 (1)rum1+遺伝子とオーレオバシジン耐性遺伝子を融合させることにより、酵母内でのRum1タンパク質の安定性検出用モニターシステムを構築した。 (2)上記遺伝子を発現している酵母細胞を変異源処理し、11株のRum1安定性に関わる変異株を取得した。 (3)これらの変異株から現在までに6クローン取得し、トランスポゾンシステムにより対応する遺伝子を特定中である。 (4)Rum1分解の阻害分子として、SUMO1に対するE2として知られているHus5、SCF複合体のSkp1に結合する分子Sgt1、その他未知の分子4つが取得された。 (5)現在これらと、Rum1のユビキチン化に関わるSCF複合体との関係を、遺伝学的生化学的に検討中である。
細胞周期制御因子の分解機構の解明とノックアウトマウスを用いた発がん研究

中山啓子, 小南欽一郎, 畠山鎮次, 北川雅敏

Offer Organization: 日本学術振興会

System: 科学研究費助成事業

Category: 特定領域研究(A)

Institution: 九州大学

1999 - 1999

More details Close

細胞周期進行に不可欠であると考えられているサイクリンEの分解機構の解明をめざし、サイクリンEのユビキチンリガーゼの同定を試みた。酵母ですでに知られている、ユビキチンリガーゼのマウス相同遺伝子を単離し、サイクリンEへの結合及び蛋白の安定性への効果を調べ、サイクリンEのユビキチンリガーゼである複合体を同定することができた。その複合体を構成するSkp2、Skp1、Cul1のノックアウトマウスを作製し、その表現型を解析することによって、生物学的機能を知るとともに、サイクリンEの分解が発がんに果たす役割を検討した。 1.サイクリンEはSCF複合体をE3とするユビキチン/プロテアゾーム系によって分解された。 2.SCF複合体を構成するF-boxタンパクはSkp2であった。 3.Skp2ノックアウトマウスは発生段階では体重が少ないこと以外に肉眼的な以上は認めないが、肝細胞や気管上皮細胞などで細胞および核の腫大が認められ、肝細胞では染色体倍数性が異常となり、4倍体、8倍体の核が多数観察された。 4.Skp2ノックアウトマウスの胎仔線維芽細胞は増殖が遅く、正常に比しアポトーシスが多く見られた。 5.Skp2ノックアウトマウスでは生後1年までで明らかな発がん傾向を認めていない。 6.Skp1及びCul1ノックアウトマウスはいずれも胎生6.5〜7.5日目に死亡する早期の胎生致死であった。 7.Skp1及びCul1ノックアウトマウスにはサイクリンEおよびβ-カテニンの蓄積が見られた。
サイクリン分解関連酵素Cdc53(Cul-1)の単離とノックアウトマウスの作製

中山啓子

Offer Organization: 日本学術振興会

System: 科学研究費助成事業

Category: 奨励研究(A)

Institution: 九州大学

1998 - 1999

More details Close

酵母ですでに知られている、ユビキチンリガーゼのマウス相同遺伝子を単離し、その生物学的機能を知るためにノックアウトマウスを作製した。 1.酵母SCF複合体のマウス相同遺伝子を、ESTdatabase上で検索し、マウスCul1のcDNA塩基配列の一部を得ることができた。 2.上記のcDNAの一部をプローブとして、マウス胸腺cDNAライブラリーをスクリーニングし、マウスCul1のcDNA上タンパクコード領域全長を得ることができた。 3.cDNA全長をプローブとし、129マウスゲノムライブラリーをスクリーニングし、9個の独立したクローンを得た。 4.上記クローンを解析することによって、マウスCul1遺伝子は少なくとも、60kbpに及ぶ大きな遺伝子であることが分かった。 5.翻訳開始コドンをコードするエクソンを含む遺伝子領域、約1kbpをネオマイシン耐性遺伝子で置換したターゲティングベタターを構築し、ES細胞にトランスフェクションし、相同組み換え体を得た。 6.相同組み換えES細胞をブラストシストにインジェクションして、キメラマウスを得、さらに交配を繰り返し、cul1ヘテロノックアウトマウスを得ることができた。 7.ヘテロノックアウトマウス同士を交配したが、ホモノックアウトマウスは産まれず、胎生致死であることが判明した。 8.Cul1ノックアウトマウスはいずれも胎生6.5〜7.5日目に死亡し、サイクリンEおよびβ-カテニンの蓄積が見られたので、致死の原因はSCF複合体によって蛋白分解を受けるべき様々な蛋白の以上蓄積に寄るものであると考えられた。
Regulation of degradation of p27Kip1, one of CDK inhibitory protein, for cancer therapy

KITAGAWA Masatoshi, NAKAYAMA Keiichi, HATAKEYAMA Shigetsugu, NAKAYAMA Keiko, HIRAI Aizan

Offer Organization: Japan Society for the Promotion of Science

System: Grants-in-Aid for Scientific Research

Category: Grant-in-Aid for Scientific Research (B)

Institution: KYUSHU UNIVERSITY

1998 - 1999

More details Close

It has been reported that the cyclin dependent kinase inhibitory proteins (CDK inhibitor) , such as p27ィイD1Kip1ィエD1 negatively regulates the G1-CDK activity in normal cells. In most cancer cells, up-regulation of G1-cyclin dependent kinase (G1-CDK) activity contributes in their malignant growth. First of all, we study the expression of p27ィイD1Kip1ィエD1 in human lung cancer using immunohistoichemistry. We found that p27ィイD1Kip1ィエD1 labeling index was decreased and p27ィイD1Kip1ィエD1 degradation activity was up-regulated in cancerious lung tissues, compared with nonneoplastic lung tissues. Therefore, it is important to study the mechanisms of p27ィイD1Kip1ィエD1 degradation. In the next study, we found that p27ィイD1Kip1ィエD1 was eliminated by two independent mechanisms, ubiquitin-mediated degradation and ubiquitin-independent processing. These tow activity was significantly high during progression from G1 to S phase. These finding suggested that inhibitor of p27ィイD1Kip1ィエD1 degradation is a potential drug for cancer therapy.
Identification and functional analysis of S phase promoting complex in mammalian cell growth

KITAGAWA Masatoshi, NAKAYAMA Keiichi, HATAKEYAMA Shigetsugu, NAKAYAMA Keiko

Offer Organization: Japan Society for the Promotion of Science

System: Grants-in-Aid for Scientific Research

Category: Grant-in-Aid for Scientific Research (B)

Institution: KYUSHU UNIVERSITY

1998 - 1999

More details Close

We study the mechanisms of growth promotion via ubiquitin-dependent proteolysis. β-catenin plays an essential role for growth regulation and colon cancer formation. Cytoplasmic β-catenin is regulated by ubiquitin-proteasome system via Wnt signaling pathway. In this study, we identified FWD1, an ubiquitin ligase for β-catenin. FWD1 formed a complex with β-catenin, axin, APC, GSK3βvia its WD40 repeats in the phosphorylation-dependent manner. Furthermore, FED1 also bound with Skp1 and Cull as an ubiquitination machinery named SCF through its F-box. The SCFィイD1FWD1ィエD1 complex ubiquitinated the β-catenin in vivo as well as in vitro. Immunohistchemical analysis indicated that transfection of FWD1 decreased the level of cytoplasmic β-catenin in SW480 colon cancer cell line. These findings suggest that FWD1 is useful for gene-therapy against colon cancer.
細胞周期調節分子の分解制御に関する研究 - Yeast three-hybrid法の確立と応用 -

畠山鎮次, 中山啓子, 北川雅敏, 中山敬一

Offer Organization: 日本学術振興会

System: 科学研究費助成事業

Category: 特定領域研究(A)

Institution: 九州大学

1998 - 1998

More details Close

細胞周期を調節するサイクリンとサイクリン依存性キナーゼ(CDK)に直接結合し、機能抑制するCDK阻害分子(CDK inhibitors;CKI)であるP27はユビキチン化を受けることにより、その発現レベルが調節されている。本研究は、p27のユビキチン化に関与するE3が存在すると仮定し、p27とUBC3と3分子複合体を形成する新たなる遺伝子を同定することを目的とした。さらに、本研究では、3分子が複合体を形成するときに、目的とする第3の遺伝子をクローニングする方法であるYeast three-hybrid法を開発した。また、p53、p27を始め、細胞周期を調節する分子(サイクリン、CKIなど)の発現が合成のみならず、分解(特にユビキチン化)により制御されていることが報告され始めている。特に、酵母における先駆的研究により、Skp1/CUl-1/E-box蛋白複合体(SCF複合体)は、CKIを含む細胞周期調節分子をはじめ、さまざまな細胞機能に関与する分子の発現調節を行っていることが報告されている。我々は、酵母におけるアナロジーを利用して、マウスにおけるSkp1、Cul-1及び複数のF-box蛋白を同定した。我々がFWD1として同定したF-box蛋白は、免疫学上重要であるNF-κBの抑制分子であるIκBαと、大腸癌に関与するβ-cateninのユビキチン化、及び分解に関与していることが明らかとなった。この結果は、酵母以外の真核細胞におけるSCF複合体によるユビキチン化介在性分解の初めての報告となる。
サイクリン依存性キナーゼ阻害分子の分解機構の解明

中山啓子, 畠山鎮次, 北川雅敏, 中山敬一

Offer Organization: 日本学術振興会

System: 科学研究費助成事業

Category: 特定領域研究(A)

Institution: 九州大学

1998 - 1998

More details Close

p27などCDKインヒビターと呼ばれる分子群は、CDK/Cyclin複合体に結合し酵素活性を阻害することから細胞周期調節に直接関わる重要な因子と考えられる。p27の蛋白量は蛋白分解に規定されていることが既に報告されていたがその分子機構はは不明であった。そこでその分解のメカニズムの解明を試み以下の点が明らかとなった。 1. 培養細胞(NIH3T3)を用い接触阻害によって細胞周期を同調させ、細胞周期によるp27の変動を観察した。p27は細胞周期依存性にG1-S移行期にもっとも強くユビキチン化された。 2. ユビキチン付加部位を特定するためp27の変異体を作製しユビキチン化を調べたところ,アミノ酸残基134番、153番、165番目のリジンをアルギニンに置換したところユビキチン化が抑制された。 3. p27をウェスタンブロットで観察すると、p27のユビキチン化と同時に22kDaの蛋白も同時に認識された。これはp27がユビキチン化以外にプロセシングを受けていることを示唆する。この反応はATP依存的であり、lactacystin、chymostatin、PMSFで抑制されたが、antipain、pepstatin、leupeptin、E64では影響を受けない。このプロセシング反応はプロテアゾームに関与しているchymotrypsin様プロテアーゼによるものと考えられた。 4. プロセシングされた22kDa蛋白(p27Δ22k)は、p27のN末端より35〜40アミノ酸が切断されたものであると予想され,この蛋白はCDK阻害活性が野生型に比べ1/100に低下していた。 5. p27Δ22kのプロセシング反応はS期に強くみられた。今回の研究の結果,p27の分解にはユビキチン化だけでなく他のプロセシング機構も関与していることが示された。ユビキチン化やプロセシングに直接関与している酵素群の同定が今後の課題である。
Analysis of sinnalling pathway of apoptosis degradation using PKCδ gene-targeting mice

NAKAYAMA Kei-ichi, HATAKEYAMA Shigetsugu, NAKAYAMA Keiko, KITAGAWA Masatoshi, CHINKAI Yoichi, OHNO Shigeo

Offer Organization: Japan Society for the Promotion of Science

System: Grants-in-Aid for Scientific Research

Category: Grant-in-Aid for Scientific Research (B)

Institution: Kyushu University

1997 - 1998

More details Close

It has been reported that protein kinase Cδ (PKCδ) a subtype of PKC family members, is cleaved by caspase resulting in conversion to activated PKCδ upon apoptosis. The aim of this research project is to elucidate the physiological function of PKCδ by generating mice deficient in PKCδ gene. We constructed the targeting vector and transfect it into embryonic stem (ES) cells to obtain the homologous recombinant of ES cells for PKCδ locus. According to the standard method to generate gene-ablated mice, PKCδ-deficient mice were produced with the mutant ES cells. The PKCδ-deficient mice were viable, healthy, and not prone to cancer development. Histlogical examinations of the PKCδ-deficient mice revealed splenomegaly due to abnormal accumulation of T and B lymphocytes. In lymph nodes, increase in the number of B cells was more apparent than that of T cells. In vitro culture experiments showed that PKCδ-deficient B cells grew faster than wild-type B cells. We concluded that PKCδ is not essential for the apoptosis during development, because of the lack of developmental abnormalities. Furthermore, apoptosis of thymovytes and embryonic fibroblasts from PKCδ-deficient mice seems unaffected. However, the extent of apoptosis in B cells appeared to be reduced, suggesting that PKCδ may play an important role in the signaling pathway of apoptosis in B cells.

Show all Show first 5

Social Activities 3

[日本国内] TAMRIC東北先進医療研究開発連携拠点

2010/01 - Present

More details Close

拠点事務局長
[日本国内] 東北大学100周年記念祭り

2007/08/25 -

More details Close

講演「大きなねずみ・小さなねずみ」
[日本国内] 東北大学オープンキャンパス全学向講演会

2007/07/30 -

More details Close

次年度入学希望者向け講演

Media Coverage 6

ＡＲＴ（東北大学大学院医学系研究科附属創生応用医学研究センター）

科学新聞

2013/07/26

Type: Newspaper, magazine

More details Close

がんコアセンターには様々な出身学部の大学院生が参画し、がん克服をめざして切磋琢磨していること、そしてその目標を共有して研究推進する経験が、研究者の育成に大きく貢献するのではないかと解説。更に具体的に進めている研究として、がん遺伝子の変異による転写やエピゲノムの変化に注目し、次世代シークエンサーを活用して網羅的な解析に取り組んでいることを紹介。
[日本国内] ラジオ番組「Crescendo」内「みんなでつくる未来の医療」コーナー

Date FM

2012/06/11

Type: Other

More details Close

「必要な新規の専門人材」というテーマで東北メディカル・メガバンク機構人材育成部門部門長として出演
[日本国内] 生命科学の先端研究拠点東北大ART がん医学コアセンター

週刊科学新聞

2012/04/20

Type: Newspaper, magazine

More details Close

がん医学コアセンターの研究内容紹介等
[日本国内] 女性医師の視点美言大義

医科同窓会会報（東京医科歯科大学お茶の水会同窓会）

2012/02/28

Type: Other

More details Close

「家庭の役割・男女の役割」というタイトルで女性医師の視点でのエッセイ
[日本国内] ヒトのメカニズム

東日本放送

2009/04/09

Type: TV or radio program

More details Close

「東北大学の新世紀」
[日本国内] 東北大学オープンキャンパス

2008/07

Type: TV or radio program

Show all Show first 5

Other 24

第１９回学際ライフサイエンスシンポジウムにおける座長
第５回山形肝炎ウイルスセミナー基調講演「次世代シークエンサーを用いた臨床研究の展開」
市民公開講演会「東北地方の復興・新生に向けて：アカデミアの果たす役割」の講演（「ゲノム科学から新しい医療に向けて」）
第１２回ＧＩリサーチフォーラム特別講演講師「臨床材料を用いた次世代シークエンサーによる新しい研究展開の可能性」
AB SCIEX プロテオミクスセミナー〜最新技術SWATHを用いたバイオマーカーの最前線〜の司会
シンポジウム『データ駆動型生命情報科学の挑戦-スーパーコンピュータ「京」と生命情報科学の接点-』での講演（「PCRから次世代シークエンサーへ：Wet研究者の奮闘」）
がん遺伝子産物RASによる広範な染色体領域にわたる転写抑制機構の解明

More details Close

がん遺伝子産物RASによる広範な染色体領域にわたる転写抑制機構の解明
第4回がん・エピゲノム研究会における座長
ユビキチンシステムの網羅的解析基盤の創出

More details Close

ユビキチンシステムの網羅的解析基盤の創出
組織特異的発がん機構の解明

More details Close

組織特異的発がん機構の解明
ユビキチンリガーゼCdc4/Fbw7の発がんに果たす役割の検討

More details Close

ユビキチンリガーゼCdc4/Fbw7の発がんに果たす役割の検討
発がんにおけるユビキチンリガーゼCdc4/Fbw7基質分子の役割

More details Close

発がんにおけるユビキチンリガーゼCdc4/Fbw7基質分子の役割
ｃ-Mycの分解異常が発がんに果たす役割

More details Close

ｃ-Mycの分解異常が発がんに果たす役割
マウス発生に関わるKPCの役割の研究

More details Close

マウス発生に関わるKPCの役割の研究
発生工学プロテオミクスを用いたPKC-δによる免疫寛容成立のメカニズムの解明

More details Close

発生工学プロテオミクスを用いたPKC-δによる免疫寛容成立のメカニズムの解明
細胞周期の再活性化による再生能力の賦活化の研究

More details Close

細胞周期の再活性化による再生能力の賦活化の研究
Mycの分解制御機構の解明

More details Close

Mycの分解制御機構の解明
神経変性疾患の発症機序の解明と遺伝子治療への応用

More details Close

神経変性疾患の発症機序の解明と遺伝子治療への応用
神経変性疾患における封入体物質のユビキチン化に関わる因子の単離同定と解析

More details Close

神経変性疾患における封入体物質のユビキチン化に関わる因子の単離同定と解析
神経再生に関する基礎的研究及び脳移植への応用

More details Close

神経再生に関する基礎的研究及び脳移植への応用
神経細胞における増殖制御機構の解明

More details Close

神経細胞における増殖制御機構の解明
リンパ球の増殖を制御する分子機構に関する研究

More details Close

リンパ球の増殖を制御する分子機構に関する研究
細胞周期制御分子の調節機構解明による新しい対癌戦略の開発

More details Close

細胞周期制御分子の調節機構解明による新しい対癌戦略の開発
PKCdノックアウトマウス作製による細胞死（アポトーシス）シグナル伝達系の研究

More details Close

PKCdノックアウトマウス作製による細胞死（アポトーシス）シグナル伝達系の研究

Show all Show first 5