Details of the Researcher

PHOTO

Nobuyuki Uozumi
Section
Graduate School of Engineering
Job title
Professor
Degree

  • 博士（工学）名古屋大学

e-Rad No.
40223515

Research History 7

  • 2007/01 - Present
    Tohoku University

  • 2004/07 - 2006/12
    Nagoya University　Bioscience and Biotechnology Center

  • 2004/10 - 2006/11
    名古屋大学高等研究院　教員（兼任）

  • 2003/06 - 2005/07
    文部科学省研究振興局　学術調査官（併任）

  • 1995/08 - 2004/06
    名古屋大学　生物分子応答研究センター　助教授

  • 1989/12 - 1995/07
    名古屋大学　工学部　助手

  • 1993/10 - 1994/10
    カリフォルニア大学　サンディエゴ校　博士研究員

Show all Show first 5

Education 4

  • 名古屋大学　大学院農学研究科　博士課程後期中退

    1988/04 - 1989/11

  • Nagoya University　Graduate School of Bioagricultural Sciences

    1986/04 - 1988/03

  • Nagoya University　School of Agricultural Sciences

    1982/04 - 1986/03

  • 博士（工学）(名古屋大学）

    1993/06 -

Research Interests 7

  • トランスポーター

  • イオンチャネル

  • バイオスティミュラント

  • 環境応答

  • 微生物

  • 植物

  • 生体膜

Research Areas 3

  • Life sciences / Applied biochemistry /

  • Environmental science/Agricultural science / Environmental agriculture /

  • Manufacturing technology (mechanical, electrical/electronic, chemical engineering) / Applied biofunctional and bioprocess engineering / Bioengineering

Papers 171

  1. Structure reveals a regulation mechanism of plant outward-rectifying K+ channel GORK by structural rearrangements in the CNBD-Ankyrin bridge. International-journal Peer-reviewed

    Taro Yamanashi, Yuki Muraoka, Tadaomi Furuta, Tsukasa Kume, Natsuko Sekido, Shunya Saito, Shota Terashima, Takeshi Yokoyama, Yoshikazu Tanaka, Atsushi Miyamoto, Kanane Sato, Tomoyuki Ito, Hikaru Nakazawa, Mitsuo Umetsu, Ellen Tanudjaja, Masaru Tsujii, Ingo Dreyer, Julian I Schroeder, Yasuhiro Ishimaru, Nobuyuki Uozumi

    Proceedings of the National Academy of Sciences of the United States of America　122　(30)　e2500070122　2025/07/29

    DOI： 10.1073/pnas.2500070122 　

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    Guard cells, which regulate stomatal apertures in plants, possess a sophisticated mechanism for regulating turgor pressure. The outward-rectifying "K+out" channel GORK, expressed in guard cells of the plant Arabidopsis thaliana, is a central component that promotes stomatal closure by releasing K+ to the extracellular space, thereby lowering turgor pressure. To date, the structural basis underlying the regulation of the K+ transport activity of GORK is unclear. Using cryo-EM, we determined the structures of the GORK outward-rectifying K+ channel with a resolution of 3.16 to 3.27 Å in five distinct conformations that differ significantly in their C-terminal cyclic nucleotide binding domain (CNBD) and ankyrin repeat (ANK) domain. The C-linker connects the transmembrane domains to the C-terminal domains, i.e., CNBD, CNBD-Ankyrin bridge, and ANK. The structural changes and interactions in the C-linker determine whether the closed state of GORK is closer to the preopen state or in a more removed state from the open state of the channel. In particular, interconversion in the short sequence within the CNBD-Ankyrin bridge plays a decisive role in this determination. This region forms an α-helix in the preopened state, while it adopts a nonhelical structure in further distant closed states. The dynamics of the cytosolic region strongly suggest that the K+ channel activity of GORK is regulated by cytosolic signaling factors during stomatal closure.

  2. Functional characterization of an additional transmembrane domain unique to TrkG and TrkH in Escherichia coli. International-journal Peer-reviewed

    Ellen Tanudjaja, Haoyu Zhang, Tadaomi Furuta, Masaru Tsujii, Yasuhiro Ishimaru, Nobuyuki Uozumi

    Bioscience, biotechnology, and biochemistry　2025/07/15

    DOI： 10.1093/bbb/zbaf101 　

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    Escherichia coli TrkG and TrkH transporters contain a unique N-terminal Domain-0 (D0). Our findings reveal that D0 supports both the function and stability of TrkG, enabling K+ and Na+ uptake, whereas it is not essential for TrkH-mediated K+ uptake. This difference can be attributed to D0 role in stabilizing polar residues within TrkG core transmembrane domains.

  3. Na+-driven pH regulation by Na+/H+ antiporters promotes photosynthetic efficiency in cyanobacteria Peer-reviewed

    Masaru Tsujii, Ayumu Kobayashi, Ayaka Kano, Kota Kera, Tomoko Takagi, Noriko Nagata, Seiji Kojima, Kouki Hikosaka, Riichi Oguchi, Kintake Sonoike, Chihiro Azai, Tomomi Inagaki, Yasuhiro Ishimaru, Nobuyuki Uozumi

    Plant Physiology　2025/01/01

    Publisher: Oxford University Press (OUP)

    DOI： 10.1093/plphys/kiae562 　

    ISSN： 0032-0889

    eISSN： 1532-2548

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    Abstract Photosynthetic organisms have developed mechanisms to regulate light reactions in response to varying light conditions. Photosynthetic electron transport leads to the formation of a ΔpH across the thylakoid membrane, which is crucial for regulating electron transport. However, other pH modulators remain to be identified, particularly in cyanobacteria. In this study, we evaluated the potential involvement of six Na+/H+ antiporters (NhaS1–NhaS6) in control of pH in the cyanobacterium Synechocystis sp. PCC 6803. Synechocystis showed a strong requirement for Na+ at high light intensities, with ΔnhaS1 and ΔnhaS2 strains unable to grow under high light conditions. We analyzed Na+ efflux–driven H+-uptake activities of NhaS1–NhaS6 in inverted membranes of Escherichia coli. Biological fractionation and immunoelectron microscopy revealed that NhaS1 localizes to both the plasma and thylakoid membranes while NhaS2 localizes to the plasma membrane. Measurement of photosynthesis activity indicated that NhaS2 promotes ATP production and electron transport from PQ to P700. Measurements of pH outside of the cells and in the cytoplasm suggested that both NhaS1 and NhaS2 are involved in plasma membrane–mediated light-dependent H+ uptake and cytoplasmic acidification. NhaS1 and NhaS2 were also found to prevent photoinhibition under high light treatment. These results indicate that H+ transport mediated by NhaS1 and NhaS2 plays a role in regulating intracellular pH and maintaining photosynthetic electron transport.

  4. Comparison of in-vacuum micro-PIXE and in-air micro-PIXE for unfixed plant sample Peer-reviewed

    Misako Miwa, Ayumi Nakatsuma, Shigeo Matsuyama, Sho Toyama, Takeshi Uchiyama, Yasuhiro Ishimaru, Nobuyuki Uozumi

    Nuclear Instruments and Methods in Physics Research Section B: Beam Interactions with Materials and Atoms　2024/09/28

    DOI： 10.1016/j.nimb.2024.165536 　

    ISSN： 0168-583X

  5. Dissecting structure and function of the monovalent cation/H + antiporters Mdm38 and Ylh47 in Saccharomyces cerevisiae Peer-reviewed

    Masaru Tsujii, Ellen Tanudjaja, Haoyu Zhang, Haruto Shimizukawa, Ayumi Konishi, Tadaomi Furuta, Yasuhiro Ishimaru, Nobuyuki Uozumi

    Journal of Bacteriology　2024/07/31

    Publisher: American Society for Microbiology

    DOI： 10.1128/jb.00182-24 　

    ISSN： 0021-9193

    eISSN： 1098-5530

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    ABSTRACT Saccharomyces cerevisiae Mdm38 and Ylh47 are homologs of the Ca 2+ /H + antiporter Letm1, a candidate gene for seizures associated with Wolf-Hirschhorn syndrome in humans. Mdm38 is important for K + /H + exchange across the inner mitochondrial membrane and contributes to membrane potential formation and mitochondrial protein translation. Ylh47 also localizes to the inner mitochondrial membrane. However, knowledge of the structures and detailed transport activities of Mdm38 and Ylh47 is limited. In this study, we conducted characterization of the ion transport activities and related structural properties of Mdm38 and Ylh47. Growth tests using Na + /H + antiporter-deficient Escherichia coli strain TO114 showed that Mdm38 and Ylh47 had Na + efflux activity. Measurement of transport activity across E. coli -inverted membranes showed that Mdm38 and Ylh47 had K + /H + , Na + /H + , and Li + /H + antiport activity, but unlike Letm1, they lacked Ca 2+ /H + antiport activity. Deletion of the ribosome-binding domain resulted in decreased Na + efflux activity in Mdm38. Structural models of Mdm38 and Ylh47 identified a highly conserved glutamic acid in the pore-forming membrane-spanning region. Replacement of this glutamic acid with alanine, a non-polar amino acid, significantly impaired the ability of Mdm38 and Ylh47 to complement the salt sensitivity of E. coli TO114. These findings not only provide important insights into the structure and function of the Letm1-Mdm38-Ylh47 antiporter family but by revealing their distinctive properties also shed light on the physiological roles of these transporters in yeast and animals. IMPORTANCE The inner membrane of mitochondria contains numerous ion transporters, including those facilitating H + transport by the electron transport chain and ATP synthase to maintain membrane potential. Letm1 in the inner membrane of mitochondria in animals functions as a Ca 2+ /H + antiporter. However, this study reveals that homologous antiporters in mitochondria of yeast, Mdm38 and Ylh47, do not transport Ca 2+ but instead are selective for K + and Na + . Additionally, the identification of conserved amino acids crucial for antiporter activity further expanded our understanding of the structure and function of the Letm1-Mdm38-Ylh47 antiporter family.

  6. Utilizing plasma-generated N2O5 gas from atmospheric air as a novel gaseous nitrogen source for plants Peer-reviewed

    Taro Yamanashi, Shouki Takeshi, Shota Sasaki, Keisuke Takashima, Toshiro Kaneko, Yasuhiro Ishimaru, Nobuyuki Uozumi

    Plant Molecular Biology　114　(2)　2024/04/08

    Publisher: Springer Science and Business Media LLC

    DOI： 10.1007/s11103-024-01438-9 　

    ISSN： 0167-4412

    eISSN： 1573-5028

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    Abstract Fixing atmospheric nitrogen for use as fertilizer is a crucial process in promoting plant growth and enhancing crop yields in agricultural production. Currently, the chemical production of nitrogen fertilizer from atmospheric N2 relies on the energy-intensive Haber–Bosch process. Therefore, developing a low-cost and easily applicable method for fixing nitrogen from the air would provide a beneficial alternative. In this study, we tested the utilization of dinitrogen pentoxide (N2O5) gas, generated from oxygen and nitrogen present in ambient air with the help of a portable plasma device, as a nitrogen source for the model plant Arabidopsis thaliana. Nitrogen-deficient plants supplied with medium treated with N2O5, were able to overcome nitrogen deficiency, similar to those provided with medium containing a conventional nitrogen source. However, prolonged direct exposure of plants to N2O5 gas adversely affected their growth. Short-time exposure of plants to N2O5 gas mitigated its toxicity and was able to support growth. Moreover, when the exposure of N2O5 and the contact with plants were physically separated, plants cultured under nitrogen deficiency were able to grow. This study shows that N2O5 gas generated from atmospheric nitrogen can be used as an effective nutrient for plants, indicating its potential to serve as an alternative nitrogen fertilization method for promoting plant growth.

  7. Instantaneous extracellular solution exchange for concurrent evaluation of membrane permeability of single cells Peer-reviewed

    Shingo Kaneko, Sugiura Hirotaka, Masaru Tsujii, Hisataka Maruyama, Nobuyuki Uozumi, Fumihito Arai

    Lab on a Chip　24　(2)　281-291　2024

    Publisher: Royal Society of Chemistry (RSC)

    DOI： 10.1039/d3lc00633f 　

    ISSN： 1473-0197

    eISSN： 1473-0189

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    The rapid osmotic stress is imposed on the microorganisms by the exchange of a locally formed droplet containing cells.

  8. An outward-rectifying plant K+ channel SPORK2 exhibits temperature-sensitive ion-transport activity Peer-reviewed

    Yuki Muraoka, Gangqiang Yang, Shintaro Munemasa, Yusuke Takeuchi, Yasuhiro Ishimaru, Yoshiyuki Murata, Nobuyuki Uozumi, Minoru Ueda

    Current Biology　33　(24)　5488-5494.e7　2023/12

    Publisher: Elsevier BV

    DOI： 10.1016/j.cub.2023.10.057 　

    ISSN： 0960-9822

  9. Integration of Microfluidic Chip and Probe with a Dual Pump System for Measurement of Single Cells Transient Response Peer-reviewed

    Xu Du, Shingo Kaneko, Hisataka Maruyama, Hirotaka Sugiura, Masaru Tsujii, Nobuyuki Uozumi, Fumihito Arai

    Micromachines　14　(6)　1210-1210　2023/06/07

    Publisher: MDPI AG

    DOI： 10.3390/mi14061210 　

    eISSN： 2072-666X

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    The integration of liquid exchange and microfluidic chips plays a critical role in the biomedical and biophysical fields as it enables the control of the extracellular environment and allows for the simultaneous stimulation and detection of single cells. In this study, we present a novel approach for measuring the transient response of single cells using a system integrated with a microfluidic chip and a probe with a dual pump. The system was composed of a probe with a dual pump system, a microfluidic chip, optical tweezers, an external manipulator, an external piezo actuator, etc. Particularly, we incorporated the probe with the dual pump to allow for high-speed liquid change, and the localized flow control enabled a low disturbance contact force detection of single cells on the chip. Using this system, we measured the transient response of the cell swelling against the osmotic shock with a very fine time resolution. To demonstrate the concept, we first designed the double-barreled pipette, which was assembled with two piezo pumps to achieve a probe with the dual pump system, allowing for simultaneous liquid injection and suction. The microfluidic chip with on-chip probes was fabricated, and the integrated force sensor was calibrated. Second, we characterized the performance of the probe with the dual pump system, and the effect of the analysis position and area of the liquid exchange time was investigated. In addition, we optimized the applied injection voltage to achieve a complete concentration change, and the average liquid exchange time was achieved at approximately 3.33 ms. Finally, we demonstrated that the force sensor was only subjected to minor disturbances during the liquid exchange. This system was utilized to measure the deformation and the reactive force of Synechocystis sp. strain PCC 6803 in osmotic shock, with an average response time of approximately 16.33 ms. This system reveals the transient response of compressed single cells under millisecond osmotic shock which has the potential to characterize the accurate physiological function of ion channels.

  10. The HKT1 Na + transporter protects plant fertility by decreasing Na + content in stamen filaments Peer-reviewed

    Takeshi Uchiyama, Shunya Saito, Taro Yamanashi, Megumi Kato, Kosuke Takebayashi, Shin Hamamoto, Masaru Tsujii, Tomoko Takagi, Noriko Nagata, Hayato Ikeda, Hidetoshi Kikunaga, Toshimi Suda, Sho Toyama, Misako Miwa, Shigeo Matsuyama, Mitsunori Seo, Tomoaki Horie, Takashi Kuromori, Mutsumi Yamagami, Yasuhiro Ishimaru, Nobuyuki Uozumi

    Science Advances　9　(22)　2023/06/02

    Publisher: American Association for the Advancement of Science (AAAS)

    DOI： 10.1126/sciadv.adg5495 　

    eISSN： 2375-2548

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    Salinity stress can greatly reduce seed production because plants are especially sensitive to salt during their reproductive stage. Here, we show that the sodium ion transporter AtHKT1;1 is specifically expressed around the phloem and xylem of the stamen in Arabidopsis thaliana to prevent a marked decrease in seed production caused by salt stress. The stamens of AtHKT1;1 mutant under salt stress overaccumulate Na + , limiting their elongation and resulting in male sterility. Specifically restricting AtHKT1;1 expression to the phloem leads to a 1.5-fold increase in the seed yield upon sodium ion stress. Expanding phloem expression of AtHKT1;1 throughout the entire plant is a promising strategy for increasing plant productivity under salinity stress.

  11. Two cyanobacterial response regulators with diguanylate cyclase activity, Rre2 and Rre8, participate in biofilm formation Peer-reviewed

    Ayumu Kobayashi, Masamune Nakamura, Masaru Tsujii, Kohei Makino, Tatsuya Nagayama, Kensuke Nakamura, Kei Nanatani, Kera Kota, Yuki Furuuchi, Shunsuke Kayamori, Tadaomi Furuta, Iwane Suzuki, Yoshihiro Hayakawa, Ellen Tanudjaja, Yasuhiro Ishimaru, Nobuyuki Uozumi

    Molecular Microbiology　119　(5)　599-611　2023/03/29

    Publisher: Wiley

    DOI： 10.1111/mmi.15057 　

    ISSN： 0950-382X

    eISSN： 1365-2958

  12. Dynamic Deformation Measurement of an Intact Single Cell via Microfluidic Chip with Integrated Liquid Exchange Peer-reviewed

    Xu Du, Di Chang, Shingo Kaneko, Hisataka Maruyama, Hirotaka Sugiura, Masaru Tsujii, Nobuyuki Uozumi, Fumihito Arai

    Engineering　2023/02

    Publisher: Elsevier BV

    DOI： 10.1016/j.eng.2022.08.020 　

    ISSN： 2095-8099

  13. Two Trk/Ktr/HKT-type potassium transporters, TrkG and TrkH, perform distinct functions in Escherichia coli K-12 Peer-reviewed

    Ellen Tanudjaja, Naomi Hoshi, Kaneyoshi Yamamoto, Kunio Ihara, Tadaomi Furuta, Masaru Tsujii, Yasuhiro Ishimaru, Nobuyuki Uozumi

    Journal of Biological Chemistry　299　(2)　102846-102846　2023/02

    Publisher: Elsevier BV

    DOI： 10.1016/j.jbc.2022.102846 　

    ISSN： 0021-9258

  14. Instantaneous extracellular solution exchange for concurrent evaluation of membrane permeability of single cells Peer-reviewed

    Shingo Kaneko, Hirotaka Sugiura, Masaru Tsujii, Hisataka Maruyama, Nobuyuki Uozumi, Fumihito Arai

    Lab on a Chip　2023

    Publisher: Royal Society of Chemistry (RSC)

    DOI： 10.1039/d3lc00633f 　

    ISSN： 1473-0197

    eISSN： 1473-0189

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    The osmotic stress imposed on microorganisms by hypotonic conditions is perceived to regulate water and solute flux via cell membranes, which are crucial for survival. Some cells that fail to...

  15. Hyper-mobile Water and Raman 2900 cm–1 Peak Band of Water Observed around Backbone Phosphates of Double Stranded DNA by High-Resolution Spectroscopies and MD Structural Feature Analysis of Water Peer-reviewed

    Makoto Suzuki, Akira Tsuchiko, Yoshiyuki Tanaka, Nobuyuki Matubayasi, George Mogami, Nobuyuki Uozumi, Satoshi Takahashi

    Journal of Physical Chemistry B　127　(1)　285-299　2022/12/27

    Publisher: American Chemical Society (ACS)

    DOI： 10.1021/acs.jpcb.2c06952 　

    ISSN： 1520-6106

    eISSN： 1520-5207

  16. Potassium transporter KUP9 participates in K+ distribution in roots and leaves under low K+ stress Invited Peer-reviewed

    Taro Yamanashi, Takeshi Uchiyama, Shunya Saito, Taiki Higashi, Hayato Ikeda, Hidetoshi Kikunaga, Mutsumi Yamagami, Yasuhiro Ishimaru, Nobuyuki Uozumi

    Stress Biology　2　(1)　2022/12/12

    Publisher: Springer Science and Business Media LLC

    DOI： 10.1007/s44154-022-00074-x 　

    eISSN： 2731-0450

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    Abstract Potassium (K) is a major essential element in plant cells, and KUP/HAK/KT-type K+ transporters participate in the absorption of K+ into roots and in the long-distance transport to above-ground parts. In Arabidopsis thaliana, KUP9 is involved in the transport of K+ and Cs+ in roots. In this study, we investigated KUP9 function in relation to the K+ status of the plant. The expression of KUP9 was upregulated in older leaves on K+-depleted medium, compared to the expression of the other 12 KUP genes in the KUP/HAK/KT family in Arabidopsis. When grown on low K+ medium, the kup9 mutant had reduced chlorophyll content in seedlings and chlorosis in older rosette leaves. Tissue-specific expression of KUP9 determined by KUP9 promoter:GUS assay depended on the K+ status of the plants: In K+ sufficient medium, KUP9 was expressed in the leaf blade towards the leaf tip, whereas in K+ depleted medium expression was mainly found in the petioles. In accordance with this, K+ accumulated in the roots of kup9 plants. The short-term 43K+ tracer measurement showed that 43K was transferred at a lower rate in roots and shoots of kup9, compared to the wild type. These data show that KUP9 participates in the distribution of K+ in leaves and K+ absorption in roots under low K+ conditions.

  17. Green Tea Catechins, (-)-Catechin Gallate, and (-)-Gallocatechin Gallate are Potent Inhibitors of ABA-Induced Stomatal Closure. International-journal Peer-reviewed

    Kanane Sato, Shunya Saito, Kohsuke Endo, Masaru Kono, Taishin Kakei, Haruka Taketa, Megumi Kato, Shin Hamamoto, Matteo Grenzi, Alex Costa, Shintaro Munemasa, Yoshiyuki Murata, Yasuhiro Ishimaru, Nobuyuki Uozumi

    Advanced Science (Weinheim, Baden-Wurttemberg, Germany)　9　(21)　e2201403　2022/05/07

    DOI： 10.1002/advs.202201403 　

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    Stomatal movement is indispensable for plant growth and survival in response to environmental stimuli. Cytosolic Ca2+ elevation plays a crucial role in ABA-induced stomatal closure during drought stress; however, to what extent the Ca2+ movement across the plasma membrane from the apoplast to the cytosol contributes to this process still needs clarification. Here the authors identify (-)-catechin gallate (CG) and (-)-gallocatechin gallate (GCG), components of green tea, as inhibitors of voltage-dependent K+ channels which regulate K+ fluxes in Arabidopsis thaliana guard cells. In Arabidopsis guard cells CG/GCG prevent ABA-induced: i) membrane depolarization; ii) activation of Ca2+ permeable cation (ICa ) channels; and iii) cytosolic Ca2+ transients. In whole Arabidopsis plants co-treatment with CG/GCG and ABA suppressed ABA-induced stomatal closure and surface temperature increase. Similar to ABA, CG/GCG inhibited stomatal closure is elicited by the elicitor peptide, flg22 but has no impact on dark-induced stomatal closure or light- and fusicoccin-induced stomatal opening, suggesting that the inhibitory effect of CG/GCG is associated with Ca2+ -related signaling pathways. This study further supports the crucial role of ICa channels of the plasma membrane in ABA-induced stomatal closure. Moreover, CG and GCG represent a new tool for the study of abiotic or biotic stress-induced signal transduction pathways.

  18. 12-Hydroxyjasmonic acid glucoside causes leaf-folding of Samanea saman through ROS accumulation. International-journal Peer-reviewed

    Gangqiang Yang, Yasuhiro Ishimaru, Shunji Hoshino, Yuki Muraoka, Nobuyuki Uozumi, Minoru Ueda

    Scientific Reports　12　(1)　7232-7232　2022/05/04

    DOI： 10.1038/s41598-022-11414-2 　

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    Foliar nyctinasty, a circadian rhythmic movement in plants, is common among leguminous plants and has been widely studied. Biological studies on nyctinasty have been conducted using Samanea saman as a model plant. It has been shown that the circadian rhythmic potassium flux from/into motor cells triggers cell shrinking/swelling to cause nyctinastic leaf-folding/opening movement in S. saman. Recently, 12-hydroxyjasmonic acid glucoside (JAG) was identified as an endogenous chemical factor causing leaf-folding of S. saman. Additionally, SPORK2 was identified as an outward-rectifying potassium channel that causes leaf-movement in the same plant. However, the molecular mechanism linking JAG and SPORK2 remains elusive. Here, we report that JAG induces leaf-folding through accumulation of reactive oxygen species in the extensor motor cells of S. saman, and this occurs independently of plant hormone signaling. Furthermore, we show that SPORK2 is indispensable for the JAG-triggered shrinkage of the motor cell. This is the first report on JAG, which is believed to be an inactivated/storage derivative of JA, acting as a bioactive metabolite in plant.

  19. Current Methods to Unravel the Functional Properties of Lysosomal Ion Channels and Transporters. International-journal Peer-reviewed

    Margherita Festa, Velia Minicozzi, Anna Boccaccio, Laura Lagostena, Antonella Gradogna, Tianwen Qi, Alex Costa, Nina Larisch, Shin Hamamoto, Emanuela Pedrazzini, Stefan Milenkovic, Joachim Scholz-Starke, Matteo Ceccarelli, Alessandro Vitale, Petra Dietrich, Nobuyuki Uozumi, Franco Gambale, Armando Carpaneto

    Cells　11　(6)　2022/03/08

    DOI： 10.3390/cells11060921 　

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    A distinct set of channels and transporters regulates the ion fluxes across the lysosomal membrane. Malfunctioning of these transport proteins and the resulting ionic imbalance is involved in various human diseases, such as lysosomal storage disorders, cancer, as well as metabolic and neurodegenerative diseases. As a consequence, these proteins have stimulated strong interest for their suitability as possible drug targets. A detailed functional characterization of many lysosomal channels and transporters is lacking, mainly due to technical difficulties in applying the standard patch-clamp technique to these small intracellular compartments. In this review, we focus on current methods used to unravel the functional properties of lysosomal ion channels and transporters, stressing their advantages and disadvantages and evaluating their fields of applicability.

  20. 放射性Naを用いた植物輸送体の機能解析

    内山 剛志, 竹林 昂亮, 加藤 恵, 鈴木 伸郎, 尹 永根, 河地 有木, 藤巻 秀, 渡辺 浩司, 池田 隼人, 菊永 英寿, 須田 利美, 遠山 翔, 三輪 美沙子, 松山 成男, 山上 睦, 石丸 泰寛, 魚住 信之

    アイソトープ・放射線研究発表会　2　129　2022

    Publisher: 公益社団法人 日本アイソトープ協会

    DOI： 10.50955/happyokai.2.0_129 　

    eISSN： 2436-4487

  21. 植物における硫黄代謝と光合成制御

    雅 辻井, 泰寛 石丸, 信之 魚住

    生化学　93　(5)　643-650　2021/10/25

    DOI： 10.14952/SEIKAGAKU.2021.930643 　

    ISSN： 0037-1017

  22. Rice amino acid transporter-like 6 (OsATL6) is involved in amino acid homeostasis by modulating the vacuolar storage of glutamine in roots. International-journal Peer-reviewed

    Saori Ogasawara, Masataka Ezaki, Ryusuke Ishida, Kuni Sueyoshi, Shunya Saito, Yuki Hiradate, Toru Kudo, Mitsuhiro Obara, Soichi Kojima, Nobuyuki Uozumi, Kentaro Tanemura, Toshihiko Hayakawa

    Plant Journal : for cell and molecular biology　107　(6)　1616-1630　2021/07/03

    DOI： 10.1111/tpj.15403 　

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    Glutamine is a product of ammonium (NH4 + ) assimilation catalyzed by glutamine synthetase (GS) and glutamate synthase (GOGAT). The growth of NH4 + -preferring paddy rice (Oryza sativa L.) depends on root NH4 + assimilation and the subsequent root-to-shoot allocation of glutamine; however, little is known about the mechanism of glutamine storage in roots. Here, using transcriptome and reverse genetics analyses, we show that the rice amino acid transporter-like 6 (OsATL6) protein exports glutamine to the root vacuoles under NH4 + -replete conditions. OsATL6 was expressed, along with OsGS1;2 and OsNADH-GOGAT1, in wild-type (WT) roots fed with sufficient NH4 Cl, and was induced by glutamine treatment. We generated two independent Tos17 retrotransposon insertion mutants showing reduced OsATL6 expression to determine the function of OsATL6. Compared with segregants lacking the Tos17 insertion, the OsATL6 knock-down mutant seedlings exhibited lower root glutamine content but higher glutamine concentration in the xylem sap and greater shoot growth under NH4 + -replete conditions. The transient expression of monomeric red fluorescent protein-fused OsATL6 in onion epidermal cells confirmed the tonoplast localization of OsATL6. When OsATL6 was expressed in Xenopus laevis oocytes, glutamine efflux from the cell into the acidic bath solution increased. Under sufficient NH4 + supply, OsATL6 transiently accumulated in sclerenchyma and pericycle cells, which are located adjacent to the Casparian strip, thus obstructing the apoplastic solute path, and in vascular parenchyma cells of WT roots before the peak accumulation of GS1;2 and NADH-GOGAT1 occurred. These findings suggest that OsATL6 temporarily stores excess glutamine, produced by NH4 + assimilation, in root vacuoles before it can be translocated to the shoot.

  23. Evaluating Young's Modulus of Single Yeast Cells Based on Compression Using an Atomic Force Microscope with a Flat Tip. International-journal Peer-reviewed

    Di Chang, Takahiro Hirate, Chihiro Uehara, Hisataka Maruyama, Nobuyuki Uozumi, Fumihito Arai

    Microscopy and microanalysis : the official journal of Microscopy Society of America, Microbeam Analysis Society, Microscopical Society of Canada　27　(2)　392-399　2021/04

    DOI： 10.1017/S1431927620024903 　

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    In this research, atomic force microscopy (AFM) with a flat tip cantilever is utilized to measure Young's modulus of a whole yeast cell (Saccharomyces cerevisiae BY4741). The results acquired from AFM are similar to those obtained using a microfluidic chip compression system. The mechanical properties of single yeast cells are important parameters which can be examined using AFM. Conventional studies apply AFM with a sharp cantilever tip to indent the cell and measure the force-indentation curve, from which Young's modulus can be calculated. However, sharp tips introduce problems because the shape variation can lead to a different result and cannot represent the stiffness of the whole cell. It can lead to a lack of broader meaning when evaluating Young's modulus of yeast cells. In this report, we confirm the differences in results obtained when measuring the compression of a poly(dimethylsiloxane) bead using a commercial sharp tip versus a unique flat tip. The flat tip effectively avoids tip-derived errors, so we use this method to compress whole yeast cells and generate a force–deformation curve. We believe our proposed method is effective for evaluating Young's modulus of whole yeast cells.

  24. Isolation of Adenosine and Cordysinin B from Anredera cordifolia that Stimulates CRE-Mediated Transcription in PC12 Cells Peer-reviewed

    Yasushi Ohizumi, Michi Kawada, Maki Kamada, Akira Nakajima, Koji Kajima, Nobuyuki Uozumi, Yasumasa Hara, Yuanqiang Guo, Masami Ishibashi

    Planta Medica International Open　8　(01)　e19-e24　2021/03

    Publisher: Georg Thieme Verlag KG

    DOI： 10.1055/a-1395-6510 　

    ISSN： 2509-9264

    eISSN： 2509-6656

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    <title>Abstract</title>Alzheimer’s disease is a typical neurodegenerative disorder, and its prevention or treatment poses great concern in advanced countries. In our survey of numerous natural resources with neurotrophic activities, we found that Anredera cordifolia improved memory impairment and increased cyclic adenosine monophosphate (AMP) response element-mediated transcription, an important step in signal transduction for memory formation. The extracts of this food were dissolved in methanol and then partitioned with three organic solvents and water, separating into n-hexane, ethyl acetate, n-butanol, and water layers. The n-butanol layer with the strongest activity on cyclic AMP-response element-dependent transcription was fractionated using silica gel column chromatography and then the activity was monitored using preparative high-performance liquid chromatography to give adenosine and cordysinin B, respectively. Both compounds showed a concentration-dependent increase in cyclic AMP-response element-mediated transcription activity. These results suggest that both adenosine and cordysinin B may participate in improving the action of A. cordifolia on memory impairment, and these actions, at least in part, result from the activation of adenosine A1, A2A, and A2B receptors.

  25. Loss of cell wall integrity genes cpxA and mrcB causes flocculation in Escherichia coli. International-journal Peer-reviewed

    Keita Sugawara, Hayato Toyoda, Mami Kimura, Shunsuke Hayasaka, Hiromi Saito, Hiroshi Kobayashi, Kunio Ihara, Tomoaki Ida, Takaaki Akaike, Eiji Ando, Mamoru Hyodo, Yoshihiro Hayakawa, Shin Hamamoto, Nobuyuki Uozumi

    Biochemical Journal　478　(1)　41-59　2021/01/15

    DOI： 10.1042/BCJ20200723 　

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    Flocculation has been recognized for hundreds of years as an important phenomenon in brewing and wastewater treatment. However, the underlying molecular mechanisms remain elusive. The lack of a distinct phenotype to differentiate between slow-growing mutants and floc-forming mutants prevents the isolation of floc-related gene by conventional mutant screening. To overcome this, we performed a two-step Escherichia coli mutant screen. The initial screen of E. coli for mutants conferring floc production during high salt treatment yielded a mutant containing point mutations in 61 genes. The following screen of the corresponding single-gene mutants identified two genes, mrcB, encoding a peptidoglycan-synthesizing enzyme and cpxA, encoding a histidine kinase of a two-component signal transduction system that contributed to salt tolerance and flocculation prevention. Both single mutants formed flocs during high salt shock, these flocs contained cytosolic proteins. ΔcpxA exhibited decreased growth with increasing floc production and addition of magnesium to ΔcpxA suppressed floc production effectively. In contrast, the growth of ΔmrcB was inconsistent under high salt conditions. In both strains, flocculation was accompanied by the release of membrane vesicles containing inner and outer membrane proteins. Of 25 histidine kinase mutants tested, ΔcpxA produced the highest amount of proteins in floc. Expression of cpxP was up-regulated by high salt in ΔcpxA, suggesting that high salinity and activation of CpxR might promote floc formation. The finding that ΔmrcB or ΔcpxA conferred floc production indicates that cell envelope stress triggered by unfavorable environmental conditions cause the initiation of flocculation in E. coli.

  26. 放射性Naを用いた植物輸送体の体内動態

    内山 剛志, 竹林 昂亮, 加藤 恵, 鈴井 伸郎, 尹 永根, 河地 有木, 藤巻 秀, 渡部 浩司, 池田 隼人, 菊永 英寿, 須田 利美, 遠山 翔, 三輪 美沙子, 松山 成男, 山上 睦, 石丸 泰寛, 魚住 信之

    アイソトープ・放射線研究発表会　1　89　2021

    Publisher: 公益社団法人 日本アイソトープ協会

    DOI： 10.50955/happyokai.1.0_89 　

    eISSN： 2436-4487

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    植物はナトリウム（Na）に曝されると生育が阻害されるため，Na+輸送体HKT1を介してNa+を輸送することで耐塩性に寄与する．これまで根の木部柔細胞におけるHKT1の耐塩性への寄与は明らかにされているが，地上部を含めた植物全体におけるHKT1の生理的役割は解明されていない。本研究では、22Naを用いてhkt1変異株とHKT1導入株におけるNa吸収および体内循環を検討した。さらに、植物におけるNaの蓄積組織の同定を行った。

  27. Analysis of Arabidopsis TPK2 and KCO3 reveals structural properties required for K+ channel function. International-journal Peer-reviewed

    Chihiro Uehara, Kota Takeda, Tatsuki Ibuki, Tadaomi Furuta, Naomi Hoshi, Ellen Tanudjaja, Nobuyuki Uozumi

    Channels (Austin, Tex.)　14　(1)　336-346　2020/12

    DOI： 10.1080/19336950.2020.1825894 　

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    Arabidopsis thaliana contains five tandem-pore domain potassium channels, TPK1-TPK5 and the related one-pore domain potassium channel, KCO3. Although KCO3 is unlikely to be an active channel, it still has a physiological role in plant cells. TPK2 is most similar to KCO3 and both are localized to the tonoplast. However, their function remains poorly understood. Here, taking advantage of the similarities between TPK2 and KCO3, we evaluated Ca2+ binding to the EF hands in TPK2, and the elements of KCO3 required for K+ channel activity. Presence of both EF-hand motifs in TPK2 resulted in Ca2+ binding, but EF1 or EF2 alone failed to interact with Ca2+. The EF hands were not required for K+ transport activity. EF1 contains two cysteines separated by two amino acids. Replacement of both cysteines with serines in TPK2 increased Ca2+ binding. We generated a two-pore domain chimeric K+ channel by replacing the missing pore region in KCO3 with a pore domain of TPK2. Alternatively, we generated two versions of simple one-pore domain K+ channels by removal of an extra region from KCO3. The chimera and one of the simple one-pore variants were functional channels. This strongly suggests that KCO3 is not a pseudogene and KCO3 retains components required for the formation of a functional K+ channel and oligomerization. Our results contribute to our understanding of the structural properties required for K+ channel activity.

  28. Hik36-Hik43 and Rre6 act as a two-component regulatory system to control cell aggregation in Synechocystis sp. PCC6803. International-journal Peer-reviewed

    Kota Kera, Yuichiro Yoshizawa, Takehiro Shigehara, Tatsuya Nagayama, Masaru Tsujii, Saeko Tochigi, Nobuyuki Uozumi

    Scientific Reports　10　(1)　19405-19405　2020/11/10

    DOI： 10.1038/s41598-020-76264-2 　

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    In response to environmental stress the model cyanobacterium, Synechocystis sp. PCC6803 can switch from a planktonic state to autoaggregation and biofilm formation. The precise mechanism of this transition remains unknown. Here we investigated the role of a candidate two-component regulatory system (TCS) in controlling morphological changes, as a way to understand the intermediate molecular steps that are part of the signaling pathway. A bacterial two-hybrid assay showed that the response regulator Rre6 formed a TCS together with a split histidine kinase consisting of Hik36 and Hik43. Individual disruption mutants displayed autoaggregation in a static culture. In contrast, unlike in the wild type, high salinity did not induce biofilm formation in Δhik36, Δhik43 and Δrre6. The expression levels of exopolysaccharide (EPS) production genes were higher in Δhik36 and Δhik43, compared with the wild type, but lower in Δrre6, suggesting that the TCS regulated EPS production in Synechocystis. Rre6 interacted physically with the motor protein PilT2, that is a component of the type IV pilus system. This interaction was enhanced in a phosphomimic version of Rre6. Taken together, Hik36-Hik43-Rre6 function as an upstream component of the pili-related signal transduction cascade and control the prevention of cell adhesion and biofilm formation.

  29. 葉緑体の光合成活性に関わるイオン輸送体の最近の知見と動向

    雅 辻井, 信之 魚住

    生化学　92　(5)　748-752　2020/10/25

    DOI： 10.14952/SEIKAGAKU.2020.920748 　

    ISSN： 0037-1017

  30. Functional characterization of multiple PAS domain-containing diguanylate cyclases in Synechocystis sp. PCC 6803. International-journal Peer-reviewed

    Ko Ishikawa, Chihiro Chubachi, Saeko Tochigi, Naomi Hoshi, Seiji Kojima, Mamoru Hyodo, Yoshihiro Hayakawa, Tadaomi Furuta, Kota Kera, Nobuyuki Uozumi

    Microbiology (Reading, England)　166　(7)　659-668　2020/07

    Publisher: Microbiology Society

    DOI： 10.1099/mic.0.000929 　

    ISSN： 1350-0872

    eISSN： 1465-2080

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    Bis-(3'-5')-cyclic dimeric guanosine monophosphate (c-di-GMP) is a second messenger known to control a variety of bacterial processes. The model cyanobacterium, Synechocystis sp. PCC 6803, has a score of genes encoding putative enzymes for c-di-GMP synthesis and degradation. However, most of them have not been functionally characterized. Here, we chose four genes in Synechocystis (dgcA-dgcD), which encode proteins with a GGDEF, diguanylate cyclase (DGC) catalytic domain and multiple Per-ARNT-Sim (PAS) conserved regulatory motifs, for detailed analysis. Purified DgcA, DgcB and DgcC were able to catalyze synthesis of c-di-GMP from two GTPs in vitro. DgcA had the highest activity, compared with DgcB and DgcC. DgcD did not show detectable activity. DgcA activity was specific for GTP and stimulated by the divalent cations, magnesium or manganese. Full activity of DgcA required the presence of the multiple PAS domains, probably because of their role in protein dimerization or stability. Synechocystis mutants carrying single deletions of dgcA-dgcD were not affected in their growth rate or biofilm production during salt stress, suggesting that there was functional redundancy in vivo. In contrast, overexpression of dgcA resulted in increased biofilm formation in the absence of salt stress. In this study, we characterize the enzymatic and physiological function of DgcA-DgcD, and propose that the PAS domains in DgcA function in maintaining the enzyme in its active form.

  31. Diverse Physiological Functions of Cation Proton Antiporters across Bacteria and Plant Cells. International-journal Peer-reviewed

    Masaru Tsujii, Ellen Tanudjaja, Nobuyuki Uozumi

    International Journal of Molecular Sciences　21　(12)　2020/06/26

    DOI： 10.3390/ijms21124566 　

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    Membrane intrinsic transport systems play an important role in maintaining ion and pH homeostasis and forming the proton motive force in the cytoplasm and cell organelles. In most organisms, cation/proton antiporters (CPAs) mediate the exchange of K+, Na+ and Ca2+ for H+ across the membrane in response to a variety of environmental stimuli. The tertiary structure of the ion selective filter and the regulatory domains of Escherichia coli CPAs have been determined and a molecular mechanism of cation exchange has been proposed. Due to symbiogenesis, CPAs localized in mitochondria and chloroplasts of eukaryotic cells resemble prokaryotic CPAs. CPAs primarily contribute to keeping cytoplasmic Na+ concentrations low and controlling pH, which promotes the detoxification of electrophiles and formation of proton motive force across the membrane. CPAs in cyanobacteria and chloroplasts are regulators of photosynthesis and are essential for adaptation to high light or osmotic stress. CPAs in organellar membranes and in the plasma membrane also participate in various intracellular signal transduction pathways. This review discusses recent advances in our understanding of the role of CPAs in cyanobacteria and plant cells.

  32. Calcium-Regulated Phosphorylation Systems Controlling Uptake and Balance of Plant Nutrients. International-journal Peer-reviewed

    Shunya Saito, Nobuyuki Uozumi

    Frontiers in Plant Science　11　44-44　2020

    DOI： 10.3389/fpls.2020.00044 　

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    Essential elements taken up from the soil and distributed throughout the whole plant play diverse roles in different tissues. Cations and anions contribute to maintenance of intracellular osmolarity and the formation of membrane potential, while nitrate, ammonium, and sulfate are incorporated into amino acids and other organic compounds. In contrast to these ion species, calcium concentrations are usually kept low in the cytosol and calcium displays unique behavior as a cytosolic signaling molecule. Various environmental stresses stimulate increases in the cytosolic calcium concentration, leading to activation of calcium-regulated protein kinases and downstream signaling pathways. In this review, we summarize the stress responsive regulation of nutrient uptake and balancing by two types of calcium-regulated phosphorylation systems: CPK and CBL-CIPK. CPK is a family of protein kinases activated by calcium. CBL is a group of calcium sensor proteins that interact with CIPK kinases, which phosphorylate their downstream targets. In Arabidopsis, quite a few ion transport systems are regulated by CPKs or CBL-CIPK complexes, including channels/transporters that mediate transport of potassium (KAT1, KAT2, GORK, AKT1, AKT2, HAK5, SPIK), sodium (SOS1), ammonium (AMT1;1, AMT1;2), nitrate and chloride (SLAC1, SLAH2, SLAH3, NRT1.1, NRT2.4, NRT2.5), and proton (AHA2, V-ATPase). CPKs and CBL-CIPKs also play a role in C/N nutrient response and in acquisition of magnesium and iron. This functional regulation by calcium-dependent phosphorylation systems ensures the growth of plants and enables them to acquire tolerance against various environmental stresses. Calcium serves as the key factor for the regulation of membrane transport systems.

  33. Calibration process for the Young's modulus of a mechanically trapped microbead measured by atomic force microscopy

    Di Chang, Takahiro Hirate, Chihiro Uehara, Nobuyuki Uozumi, Fumihito Arai

    MHS 2019 - 30th 2019 International Symposium on Micro-NanoMechatronics and Human Science　2019/12/01

    Publisher: Institute of Electrical and Electronics Engineers Inc.

    DOI： 10.1109/MHS48134.2019.9249340 　

  34. DAY-LENGTH-DEPENDENT DELAYED-GREENING1, the Arabidopsis Homolog of the Cyanobacterial H+-Extrusion Protein, Is Essential for Chloroplast pH Regulation and Optimization of Non-Photochemical Quenching. Peer-reviewed

    Kyohei Harada, Takatoshi Arizono, Ryoichi Sato, Mai Duy Luu Trinh, Akira Hashimoto, Masaru Kono, Masaru Tsujii, Nobuyuki Uozumi, Shinichi Takaichi, Shinji Masuda

    Plant & Cell Physiology　60　(12)　2660-2671　2019/12/01

    DOI： 10.1093/pcp/pcz203 　

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    Plants convert solar energy into chemical energy through photosynthesis, which supports almost all life activities on earth. Because the intensity and quality of sunlight can change dramatically throughout the day, various regulatory mechanisms help plants adjust their photosynthetic output accordingly, including the regulation of light energy accumulation to prevent the generation of damaging reactive oxygen species. Non-photochemical quenching (NPQ) is a regulatory mechanism that dissipates excess light energy, but how it is regulated is not fully elucidated. In this study, we report a new NPQ-regulatory protein named Day-Length-dependent Delayed-Greening1 (DLDG1). The Arabidopsis DLDG1 associates with the chloroplast envelope membrane, and the dldg1 mutant had a large NPQ value compared with wild type. The mutant also had a pale-green phenotype in developing leaves but only under continuous light; this phenotype was not observed when dldg1 was cultured in the dark for ≥8 h/d. DLDG1 is a homolog of the plasma membrane-localizing cyanobacterial proton-extrusion-protein A that is required for light-induced H+ extrusion and also shows similarity in its amino-acid sequence to that of Ycf10 encoded in the plastid genome. Arabidopsis DLDG1 enhances the growth-retardation phenotype of the Escherichia coli K+/H+ antiporter mutant, and the everted membrane vesicles of the E. coli expressing DLDG1 show the K+/H+ antiport activity. Our findings suggest that DLDG1 functionally interacts with Ycf10 to control H+ homeostasis in chloroplasts, which is important for the light-acclimation response, by optimizing the extent of NPQ.

  35. The mechanosensitive channel YbdG from Escherichia coli has a role in adaptation to osmotic up-shock. International-journal Peer-reviewed

    Shun Amemiya, Hayato Toyoda, Mami Kimura, Hiromi Saito, Hiroshi Kobayashi, Kunio Ihara, Kiyoto Kamagata, Ryuji Kawabata, Setsu Kato, Yutaka Nakashimada, Tadaomi Furuta, Shin Hamamoto, Nobuyuki Uozumi

    The Journal of biological chemistry　294　(33)　12281-12292　2019/08/16

    DOI： 10.1074/jbc.RA118.007340 　

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    Mechanosensitive channels play an important role in the adaptation of cells to hypo-osmotic shock. Among members of this channel family in Escherichia coli, the exact function and physiological role of the mechanosensitive channel homolog YbdG remain unclear. Characterization of YbdG's physiological role has been hampered by its lack of measurable transport activity. Using a nitrosoguanidine mutagenesis-aided screen in combination with next-generation sequencing, here we isolated a mutant with a point mutation in ybdG This mutation (resulting in a I167T change) conferred sensitivity to high osmotic stress, and the mutant cells differed from WT cells in morphology during hyperosmotic stress at alkaline pH. Interestingly, unlike the cells containing the I167T variant, a null-ybdG mutant did not exhibit this sensitivity and phenotype. Although I167T was located near the putative ion-conducting pore in a transmembrane region of YbdG, no change in ion channel activities of YbdG-I167T was detected. Of note, introduction of the WT C-terminal cytosolic region of YbdG into the I167T variant complemented the osmo-sensitive phenotype. Co-precipitation of proteins interacting with the C-terminal YbdG region led to the isolation of HldD and FbaA, whose overexpression in cells containing the YbdG-I167T variant partially rescued the osmo-sensitive phenotype. This study indicates that YbdG functions as a component of a mechanosensing system that transmits signals triggered by external osmotic changes to intracellular factors. The cellular role of YbdG uncovered here goes beyond its predicted function as an ion or solute transport protein.

  36. Evidence for potassium transport activity of Arabidopsis KEA1-KEA6. International-journal Peer-reviewed

    Masaru Tsujii, Kota Kera, Shin Hamamoto, Takashi Kuromori, Toshiharu Shikanai, Nobuyuki Uozumi

    Scientific reports　9　(1)　10040-10040　2019/07/11

    DOI： 10.1038/s41598-019-46463-7 　

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    Arabidopsis thaliana contains the putative K+ efflux transporters KEA1-KEA6, similar to KefB and KefC of Escherichia coli. KEA1-KEA3 are involved in the regulation of photosynthetic electron transport and chloroplast development. KEA4-KEA6 mediate pH regulation of the endomembrane network during salinity stress. However, the ion transport activities of KEA1-KEA6 have not been directly characterized. In this study, we used an E. coli expression system to examine KEA activity. KEA1-KEA3 and KEA5 showed bi-directional K+ transport activity, whereas KEA4 and KEA6 functioned as a K+ uptake system. The thylakoid membrane-localized Na+/H+ antiporter NhaS3 from the model cyanobacterium Synechocystis is the closest homolog of KEA3. Changing the putative Na+/H+ selective site of KEA3 (Gln-Asp) to that of NhaS3 (Asp-Asp) did not alter the ion selectivity without loss of K+ transport activity. The first residue in the conserved motif was not a determinant for K+ or Na+ selectivity. Deletion of the possible nucleotide-binding KTN domain from KEA3 lowered K+ transport activity, indicating that the KTN domain was important for this function. The KEA3-G422R mutation discovered in the Arabidopsis dpgr mutant increased K+ transport activity, consistent with the mutant phenotype. These results indicate that Arabidopsis KEA1-KEA6 act as K+ transport systems, and support the interpretation that KEA3 promotes dissipation of ΔpH in the thylakoid membrane.

  37. Cesium Inhibits Plant Growth Primarily Through Reduction of Potassium Influx and Accumulation in Arabidopsis. Peer-reviewed

    Eri Adams, Takae Miyazaki, Shunya Saito, Nobuyuki Uozumi, Ryoung Shin

    Plant & cell physiology　60　(1)　63-76　2019/01/01

    DOI： 10.1093/pcp/pcy188 　

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    Cesium (Cs+) is known to compete with the macronutrient potassium (K+) inside and outside of plants and to inhibit plant growth at high concentrations. However, the detailed molecular mechanisms of how Cs+ exerts its deleterious effects on K+ accumulation in plants are not fully elucidated. Here, we show that mutation in a member of the major K+ channel AKT1-KC1 complex renders Arabidopsis thaliana hypersensitive to Cs+. Higher severity of the phenotype and K+ loss were observed for these mutants in response to Cs+ than to K+ deficiency. Electrophysiological analysis demonstrated that Cs+, but not sodium, rubidium or ammonium, specifically inhibited K+ influx through the AKT1-KC1 complex. In contrast, Cs+ did not inhibit K+ efflux through the homomeric AKT1 channel that occurs in the absence of KC1, leading to a vast loss of K+. Our observation suggests that reduced K+ accumulation due to blockage/competition in AKT1 and other K+ transporters/channels by Cs+ plays a major role in plant growth retardation. This report describes the mechanical role of Cs+ in K+ accumulation, and in turn in plant performance, providing actual evidence at the plant level for what has long been believed, i.e. K+ channels are, therefore AKT1 is, 'blocked' by Cs+.

  38. Guard cell membrane anion transport sGuard cell membrane anion transport systems and their regulatory components: An elaborate mechanism controlling stress-induced stomatal closureystems and their regulatory components: An elaborate mechanism controlling Peer-reviewed

    Saito, S, Uozumi, N

    Plants　8　(1)　9　2019

  39. Mechanical Characterization of a Single Yeast Cell Using a Robot Integrated Microfluidic Chip

    Di Chang, Shinya Sakuma, Chihiro Uehara, Nobuyuki Uozumi, Fumihito Arai

    MHS 2018 - 2018 29th International Symposium on Micro-NanoMechatronics and Human Science　2018/12/01

    Publisher: Institute of Electrical and Electronics Engineers Inc.

    DOI： 10.1109/MHS.2018.8886914 　

  40. Reduction of Spermidine Content Resulting from Inactivation of Two Arginine Decarboxylases Increases Biofilm Formation in Synechocystis sp. Strain PCC 6803. Peer-reviewed

    Kera K, Nagayama T, Nanatani K, Saeki-Yamoto C, Tominaga A, Souma S, Miura N, Takeda K, Kayamori S, Ando E, Higashi K, Igarashi K, Uozumi N

    Journal of Bacteriology　200　(9)　E00664-17　2018/09

  41. N-myristoylation and S-acylation are common modifications of Ca2+-regulated Arabidopsis kinases and are required for activation of the SLAC1 anion channel Peer-reviewed

    Shunya Saito, Shin Hamamoto, Koko Moriya, Aiko Matsuura, Yoko Sato, Jun Muto, Hiroto Noguchi, Seiji Yamauchi, Yuzuru Tozawa, Minoru Ueda, Kenji Hashimoto, Philipp Köster, Qiuyan Dong, Katrin Held, Jörg Kudla, Toshihiko Utsumi, Nobuyuki Uozumi

    New Phytologist　218　(4)　1504-1521　2018/06/01

    Publisher: Blackwell Publishing Ltd

    DOI： 10.1111/nph.15053 　

    ISSN： 1469-8137 0028-646X

  42. Measurement of the mechanical properties of single: Synechocystis sp. strain PCC6803 cells in different osmotic concentrations using a robot-integrated microfluidic chip Peer-reviewed

    Di Chang, Shinya Sakuma, Kota Kera, Nobuyuki Uozumi, Fumihito Arai

    Lab on a Chip　18　(8)　1241-1249　2018

    Publisher: Royal Society of Chemistry

    DOI： 10.1039/c7lc01245d 　

    ISSN： 1473-0189 1473-0197

    eISSN： 1473-0189

  43. Identification and characterization of compounds that affect stomatal movements Peer-reviewed

    Toh, S, Inoue, S, Toda Y, Yuki, T, Suzuki, K, Hamamoto, S, Fukatsu, K, Aoki, S, Uchida, M, Asai, E, Uozumi, N, Ayato S, Kinoshita, T

    Plant & Cell Physiology　59　1568-1580　2018

  44. Ion Channels Regulate Nyctinastic Leaf Opening in Samanea saman Peer-reviewed

    Takaya Oikawa, Yasuhiro Ishimaru, Shintaro Munemasa, Yusuke Takeuchi, Kento Washiyama, Shin Hamamoto, Nobuyuki Yoshikawa, Yoshiyuki Mutara, Nobuyuki Uozumi, Minoru Ueda

    Current Biology　28　2230-2238　2018

    Publisher: Cell Press

    DOI： 10.1016/j.cub.2018.05.042 　

    ISSN： 0960-9822

  45. In vitro and in vivo characterization of modulation of the vacuolar cation channel TRPY1 from Saccharomyces cerevisiae Peer-reviewed

    Hamamoto, S, Mori, Y, Yabe, I, Uozumi,N

    FEBS Journal　285　1146-1161　2018

  46. Identification of regions responsible for the function of the plant K+ channels KAT1 and AKT2 in Saccharomyces cerevisiae and Xenopus laevis oocytes Peer-reviewed

    Shunya Saito, Naomi Hoshi, Lalu Zulkifli, Sri Widyastuti, Shinobu Goshima, Ingo Dreyer, Nobuyuki Uozumi

    Channels　11　(6)　510-516　2017/11/02

    Publisher: Taylor and Francis Inc.

    DOI： 10.1080/19336950.2017.1372066 　

    ISSN： 1933-6969 1933-6950

  47. Dimerization of GTR1 regulates their plasma membrane localization Peer-reviewed

    Yasuhiro Ishimaru, Kento Washiyama, Takaya Oikawa, Shin Hamamoto, Nobuyuki Uozumi, Minoru Ueda

    Plant Signaling and Behavior　12　(6)　e1334749　2017/06/03

    Publisher: Taylor and Francis Inc.

    DOI： 10.1080/15592324.2017.1334749 　

    ISSN： 1559-2324 1559-2316

  48. Probing native metal ion association sites through quenching of fluorophores in the nucleotide-binding domains of the ABC transporter MsbA Peer-reviewed

    Daiki Tatsumi, Kei Nanatani, Yuto Koike, Kiyoto Kamagata, Satoshi Takahashi, Ayumu Konno, Tadaomi Furuta, Minoru Sakurai, Nobuyuki Uozumi

    Biochemical Journal　474　1993-2007　2017/06

    DOI： 10.1042/BCJ20161051 　

    ISSN： 0264-6021

    eISSN： 1470-8728

  49. Kup-mediated Cs+ uptake and Kdp-driven K+ uptake coordinate to promote cell growth during excess Cs+ conditions in Escherichia coli Peer-reviewed

    Ellen Tanudjaja, Naomi Hoshi, Yi-Hsin Su, Shin Hamamoto, Nobuyuki Uozumi

    SCIENTIFIC REPORTS　7　2122　2017/05

    DOI： 10.1038/s41598-017-02164-7 　

    ISSN： 2045-2322

  50. GTR1 is a jasmonic acid and jasmonoyl-L-isoleucine transporter in Arabidopsis thaliana Peer-reviewed

    Yasuhiro Ishimaru, Takaya Oikawa, Takeshi Suzuki, Syohei Takeishi, Hideyuki Matsuura, Kosaku Takahashi, Shin Hamamoto, Nobuyuki Uozumi, Takafumi Shimizu, Mitsunori Seo, Hiroyuki Ohta, Minoru Ueda

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY　81　(2)　249-255　2017/02

    DOI： 10.1080/09168451.2016.1246174 　

    ISSN： 0916-8451

    eISSN： 1347-6947

  51. Mechanical Characterization of a Single Synechocystis sp. PCC 6803 Cell in Different Osmolarity Solutions

    Chang Di, Sakuma Shinya, Kera Kota, Uozumi Nobuyuki, Arai Fumihito

    2017 INTERNATIONAL SYMPOSIUM ON MICRO-NANOMECHATRONICS AND HUMAN SCIENCE (MHS)　.　2017

    ISSN： 2474-378X

  52. The topogenic function of S4 promotes membrane insertion of the voltage-sensor domain in the KvAP channel Peer-reviewed

    Eriko Mishima, Yoko Sato, Kei Nanatani, Naomi Hoshi, Jong-Kook Lee, Nina Schiller, Gunnar von Heijne, Masao Sakaguchi, Nobuyuki Uozumi

    BIOCHEMICAL JOURNAL　473　4361-4372　2016/12

    DOI： 10.1042/BCJ20160746 　

    ISSN： 0264-6021

    eISSN： 1470-8728

  53. Nerve growth factor enhances the CRE-dependent transcriptional activity activated by nobiletin in PC12 cells Peer-reviewed

    Jiro Takito, Junko Kimura, Koji Kajima, Nobuyuki Uozumi, Makoto Watanabe, Akihito Yokosuka, Yoshihiro Mimaki, Masanori Nakamura, Yasushi Ohizumi

    CANADIAN JOURNAL OF PHYSIOLOGY AND PHARMACOLOGY　94　(7)　728-733　2016/07

    DOI： 10.1139/cjpp-2015-0394 　

    ISSN： 0008-4212

    eISSN： 1205-7541

  54. Involvement of Potassium Transport Systems in the Response of Synechocystis PCC 6803 Cyanobacteria to External pH Change, High-Intensity Light Stress and Heavy Metal Stress Peer-reviewed

    Vanessa Checchetto, Anna Segalla, Yuki Sato, Elisabetta Bergantino, Ildiko Szabo, Nobuyuki Uozumi

    PLANT AND CELL PHYSIOLOGY　57　(4)　862-877　2016/04

    DOI： 10.1093/pcp/pcw032 　

    ISSN： 0032-0781

    eISSN： 1471-9053

  55. Ion Channels in Plant Bioenergetic Organelles, Chloroplasts and Mitochondria: From Molecular Identification to Function Peer-reviewed

    Luca Carraretto, Enrico Teardo, Vanessa Checchetto, Giovanni Finazzi, Nobuyuki Uozumi, Ildiko Szabo

    MOLECULAR PLANT　9　(3)　371-395　2016/03

    DOI： 10.1016/j.molp.2015.12.004 　

    ISSN： 1674-2052

    eISSN： 1752-9867

  56. Limonene enhances the cAMP response element (CRE)-dependent transcriptional activity activated via adenosine A2A receptor in a neural-crest derived cell line, PC-12 Peer-reviewed

    Takito, J, Ota, M, Oba, C, Fujiwara, H, Murata, K, Yamaguchi, K, Uozumi, N, Nakamura, M, Inomata, H, Ohizumi, Y

    Planta Medica International Open　3　(3)　e60-e62　2016

    DOI： 10.1055/s-0042-118778 　

  57. The Phytosiderophore Efflux Transporter TOM2 Is Involved in Metal Transport in Rice Peer-reviewed

    Tomoko Nozoye, Seiji Nagasaka, Takanori Kobayashi, Yuki Sato, Nobuyuki Uozumi, Hiromi Nakanishi, Naoko K. Nishizawa

    JOURNAL OF BIOLOGICAL CHEMISTRY　290　(46)　27688-27699　2015/11

    DOI： 10.1074/jbc.M114.635193 　

    eISSN： 1083-351X

  58. Iron deficiency regulated OsOPT7 is essential for iron homeostasis in rice Peer-reviewed

    Khurram Bashir, Yasuhiro Ishimaru, Reiko Nakanishi Itai, Takeshi Senoura, Michiko Takahashi, Gynheung An, Takaya Oikawa, Minoru Ueda, Aiko Sato, Nobuyuki Uozumi, Hiromi Nakanishi, Naoko K. Nishizawa

    PLANT MOLECULAR BIOLOGY　88　(1-2)　165-176　2015/05

    DOI： 10.1007/s11103-015-0315-0 　

    ISSN： 0167-4412

    eISSN： 1573-5028

  59. HKT transporters mediate salt stress resistance in plants: from structure and function to the field Peer-reviewed

    Shin Hamamoto, Tomoaki Horie, Felix Hauser, Ulrich Deinlein, Julian I. Schroeder, Nobuyuki Uozumi

    CURRENT OPINION IN BIOTECHNOLOGY　32　113-120　2015/04

    DOI： 10.1016/j.copbio.2014.11.025 　

    ISSN： 0958-1669

    eISSN： 1879-0429

  60. The jasmonate-responsive GTR1 transporter is required for gibberellin-mediated stamen development in Arabidopsis Peer-reviewed

    Hikaru Saito, Takaya Oikawa, Shin Hamamoto, Yasuhiro Ishimaru, Miyu Kanamori-Sato, Yuko Sasaki-Sekimoto, Tomoya Utsumi, Jing Chen, Yuri Kanno, Shinji Masuda, Yuji Kamiya, Mitsunori Seo, Nobuyuki Uozumi, Minoru Ueda, Hiroyuki Ohta

    NATURE COMMUNICATIONS　6　6095　2015/02

    DOI： 10.1038/ncomms7095 　

    ISSN： 2041-1723

  61. Comparative Analysis of kdp and ktr Mutants Reveals Distinct Roles of the Potassium Transporters in the Model Cyanobacterium Synechocystis sp Strain PCC 6803 Peer-reviewed

    Kei Nanatani, Toshiaki Shijuku, Yousuke Takano, Lalu Zulkifli, Tomoko Yamazaki, Akira Tominaga, Satoshi Souma, Kiyoshi Onai, Megumi Morishita, Masahiro Ishiura, Martin Hagemann, Iwane Suzuki, Hisataka Maruyama, Fumihito Arai, Nobuyuki Uozumi

    JOURNAL OF BACTERIOLOGY　197　(4)　676-687　2015/02

    DOI： 10.1128/JB.02276-14 　

    ISSN： 0021-9193

    eISSN： 1098-5530

  62. A Cell-Free Translocation System Using Extracts of Cultured Insect Cells to Yield Functional Membrane Proteins Peer-reviewed

    Toru Ezure, Kei Nanatani, Yoko Sato, Satomi Suzuki, Keishi Aizawa, Satoshi Souma, Masaaki Ito, Takahiro Hohsaka, Gunnar von Heijine, Toshihiko Utsumi, Keietsu Abe, Eiji Ando, Nobuyuki Uozumi

    PLOS ONE　9　(12)　e112874　2014/12

    DOI： 10.1371/journal.pone.0112874 　

    ISSN： 1932-6203

  63. The phosphoinositide PI(3,5)P-2 mediates activation of mammalian but not plant TPC proteins: functional expression of endolysosomal channels in yeast and plant cells Peer-reviewed

    Anna Boccaccio, Joachim Scholz-Starke, Shin Hamamoto, Nina Larisch, Margherita Festa, Paul Vijay Kanth Gutla, Alex Costa, Petra Dietrich, Nobuyuki Uozumi, Armando Carpaneto

    CELLULAR AND MOLECULAR LIFE SCIENCES　71　(21)　4275-4283　2014/11

    DOI： 10.1007/s00018-014-1623-2 　

    ISSN： 1420-682X

    eISSN： 1420-9071

  64. Molecular cloning and expression analysis of a gene encoding KUP/HAK/KT-type potassium uptake transporter from Cryptomeria japonica Peer-reviewed

    Yoshihiro Hosoo, Yukiya Kimura, Kei Nanatani, Nobuyuki Uozumi

    TREES-STRUCTURE AND FUNCTION　28　(5)　1527-1537　2014/10

    DOI： 10.1007/s00468-014-1059-1 　

    ISSN： 0931-1890

    eISSN： 1432-2285

  65. Defining membrane spanning domains and crucial membrane-localized acidic amino acid residues for K+ transport of a Kup/HAK/KT-type Escherichia coli potassium transporter Peer-reviewed

    Yoko Sato, Kei Nanatani, Shin Hamamoto, Makoto Shimizu, Miho Takahashi, Mayumi Tabuchi-Kobayashi, Akifumi Mizutani, Julian I. Schroeder, Satoshi Souma, Nobuyuki Uozumi

    JOURNAL OF BIOCHEMISTRY　155　(5)　315-323　2014/05

    DOI： 10.1093/jb/mvu007 　

    ISSN： 0021-924X

    eISSN： 1756-2651

  66. Organelle-localized potassium transport systems in plants Peer-reviewed

    Shin Hamamoto, Nobuyuki Uozumi

    JOURNAL OF PLANT PHYSIOLOGY　171　(9)　743-747　2014/05

    DOI： 10.1016/j.jplph.2013.09.022 　

    ISSN： 0176-1617

    eISSN： 1618-1328

  67. The wheat chloroplastic proteome Peer-reviewed

    Abu Hena Mostafa Kamal, Kun Cho, Jong-Soon Choi, Kwang-Hee Bae, Setsuko Komatsu, Nobuyuki Uozumi, Sun Hee Woo

    JOURNAL OF PROTEOMICS　93　326-342　2013/11

    DOI： 10.1016/j.jprot.2013.03.009 　

    ISSN： 1874-3919

    eISSN： 1876-7737

  68. Characterization of the role of a mechanosensitive channel in osmotic down shock adaptation in Synechocystis sp PCC 6803 Peer-reviewed

    Kei Nanatani, Toshiaki Shijuku, Masaro Akai, Yoshinori Yukutake, Masato Yasui, Shin Hamamoto, Kiyoshi Onai, Megumi Morishita, Masahiro Ishiura, Nobuyuki Uozumi

    CHANNELS　7　(4)　238-242　2013/07

    DOI： 10.4161/chan.25350 　

    ISSN： 1933-6950

    eISSN： 1933-6969

  69. Molecular bases of multimodal regulation of a fungal transient receptor potential (TRP) channel Peer-reviewed

    Makoto Ihara, Shin Hamamoto, Yohei Miyanoiri, Mitsuhiro Takeda, Masatsune Kainosho, Isamu Yabe, Nobuyuki Uozumi, Atsuko Yamashita

    Journal of Biological Chemistry　288　(21)　15303-15317　2013/05/24

    DOI： 10.1074/jbc.M112.434795 　

    ISSN： 0021-9258 1083-351X

  70. Sodium transport system in plant cells Invited Peer-reviewed

    Toshio Yamaguchi, Shin Hamamoto, Nobuyuki Uozumi

    Frontiers in Plant Science　4　2013

    Publisher: Frontiers Media SA

    DOI： 10.3389/fpls.2013.00410 　

    eISSN： 1664-462X

  71. A simple fed-batch method for transcription and insect cell-free translation Peer-reviewed

    Yoko Sato, Keishi Aizawa, Toru Ezure, Eiji Ando, Nobuyuki Uozumi

    JOURNAL OF BIOSCIENCE AND BIOENGINEERING　114　(6)　677-679　2012/12

    DOI： 10.1016/j.jbiosc.2012.06.015 　

    ISSN： 1389-1723

  72. Aquaporin AqpZ Is Involved in Cell Volume Regulation and Sensitivity to Osmotic Stress in Synechocystis sp Strain PCC 6803 Peer-reviewed

    Masaro Akai, Kiyoshi Onai, Megumi Morishita, Hiroyuki Mino, Toshiaki Shijuku, Hisataka Maruyama, Fumihito Arai, Shigeru Itoh, Akihiro Hazama, Vanessa Checchetto, Ildiko Szabo, Yoshinori Yukutake, Makoto Suematsu, Masato Yasui, Masahiro Ishiura, Nobuyuki Uozumi

    JOURNAL OF BACTERIOLOGY　194　(24)　6828-6836　2012/12

    DOI： 10.1128/JB.01665-12 　

    ISSN： 0021-9193

  73. Thylakoid potassium channel is required for efficient photosynthesis in cyanobacteria Peer-reviewed

    Vanessa Checchetto, Anna Segalla, Guillaume Allorent, Nicoletta La Rocca, Luigi Leanza, Giorgio Mario Giacometti, Nobuyuki Uozumi, Giovanni Finazzi, Elisabetta Bergantino, Ildiko Szabo

    PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA　109　(27)　11043-11048　2012/07

    DOI： 10.1073/pnas.1205960109 　

    ISSN： 0027-8424

  74. Towards an understanding of wheat chloroplasts: a methodical investigation of thylakoid proteome Peer-reviewed

    Abu Hena Mostafa Kamal, Kun Cho, Setsuko Komatsu, Nobuyuki Uozumi, Jong-Soon Choi, Sun Hee Woo

    MOLECULAR BIOLOGY REPORTS　39　(5)　5069-5083　2012/05

    DOI： 10.1007/s11033-011-1302-4 　

    ISSN： 0301-4851

    eISSN： 1573-4978

  75. Rice phenolics efflux transporter 2 (PEZ2) plays an important role in solubilizing apoplasmic iron Peer-reviewed

    Khurram Bashir, Yasuhiro Ishimaru, Hugo Shimo, Yusuke Kakei, Takeshi Senoura, Ryuichi Takahashi, Yutaka Sato, Yuki Sato, Nobuyuki Uozumi, Hiromi Nakanishi, Naoko K. Nishizawa

    SOIL SCIENCE AND PLANT NUTRITION　57　(6)　803-812　2011/12

    DOI： 10.1080/00380768.2011.637305 　

    ISSN： 0038-0768

  76. Uniquely evolved plant ion channels Invited Peer-reviewed

    Nobuyuki Uozumi

    FEBS JOURNAL　278　(22)　4261-4261　2011/11

    DOI： 10.1111/j.1742-4658.2011.08372.x 　

    ISSN： 1742-464X

  77. Potassium channels in plant cells Invited Peer-reviewed

    Ingo Dreyer, Nobuyuki Uozumi

    FEBS JOURNAL　278　(22)　4293-4303　2011/11

    DOI： 10.1111/j.1742-4658.2011.08371.x 　

    ISSN： 1742-464X

    eISSN： 1742-4658

  78. Plant-Specific Cation/H+ Exchanger 17 and Its Homologs Are Endomembrane K+ Transporters with Roles in Protein Sorting Peer-reviewed

    Salil Chanroj, Yongxian Lu, Senthilkumar Padmanaban, Kei Nanatani, Nobuyuki Uozumi, Rajini Rao, Heven Sze

    JOURNAL OF BIOLOGICAL CHEMISTRY　286　(39)　33931-33941　2011/09

    DOI： 10.1074/jbc.M111.252650 　

    ISSN： 0021-9258

  79. Plasma Membrane Aquaporin AqpZ Protein Is Essential for Glucose Metabolism during Photomixotrophic Growth of Synechocystis sp PCC 6803 Peer-reviewed

    Masaro Akai, Kiyoshi Onai, Miyako Kusano, Mayuko Sato, Henning Redestig, Kiminori Toyooka, Megumi Morishita, Hiroshi Miyake, Akihiro Hazama, Vanessa Checchetto, Ildiko Szabo, Ken Matsuoka, Kazuki Saito, Masato Yasui, Masahiro Ishiura, Nobuyuki Uozumi

    JOURNAL OF BIOLOGICAL CHEMISTRY　286　(28)　25224-25235　2011/07

    DOI： 10.1074/jbc.M111.236380 　

    ISSN： 0021-9258

  80. A Rice Phenolic Efflux Transporter Is Essential for Solubilizing Precipitated Apoplasmic Iron in the Plant Stele Peer-reviewed

    Yasuhiro Ishimaru, Yusuke Kakei, Hugo Shimo, Khurram Bashir, Yutaka Sato, Yuki Sato, Nobuyuki Uozumi, Hiromi Nakanishi, Naoko K. Nishizawa

    JOURNAL OF BIOLOGICAL CHEMISTRY　286　(28)　24649-24655　2011/07

    DOI： 10.1074/jbc.M111.221168 　

    ISSN： 0021-9258

  81. 12-Hydroxyjasmonic Acid Glucoside Is a COI1-JAZ-Independent Activator of Leaf-Closing Movement in Samanea saman Peer-reviewed

    Yoko Nakamura, Axel Mithoefer, Erich Kombrink, Wilhelm Boland, Shin Hamamoto, Nobuyuki Uozumi, Kentaro Tohma, Minoru Ueda

    PLANT PHYSIOLOGY　155　(3)　1226-1236　2011/03

    DOI： 10.1104/pp.110.168617 　

    ISSN： 0032-0889

  82. Phytosiderophore Efflux Transporters Are Crucial for Iron Acquisition in Graminaceous Plants Peer-reviewed

    Tomoko Nozoye, Seiji Nagasaka, Takanori Kobayashi, Michiko Takahashi, Yuki Sato, Yoko Sato, Nobuyuki Uozumi, Hiromi Nakanishi, Naoko K. Nishizawa

    JOURNAL OF BIOLOGICAL CHEMISTRY　286　(7)　5446-5454　2011/02

    DOI： 10.1074/jbc.M110.180026 　

    eISSN： 1083-351X

  83. Pollen Tubes Lacking a Pair of K+ Transporters Fail to Target Ovules in Arabidopsis Peer-reviewed

    Yongxian Lu, Salil Chanroj, Lalu Zulkifli, Mark A. Johnson, Nobuyuki Uozumi, Alice Cheung, Heven Sze

    PLANT CELL　23　(1)　81-93　2011/01

    DOI： 10.1105/tpc.110.080499 　

    ISSN： 1040-4651

    eISSN： 1532-298X

  84. Arabidopsis CHX16-20 are endomembrane cation transporters with distinct activities and emerging role in protein sorting. Peer-reviewed

    Chanroj, S, Lu, Y, Padmanaban, S, Nanatani, K, Uozumi, N, Rao R, Sze. H

    J. Biol. Chem.　2011

  85. The KtrA and KtrE Subunits Are Required for Na+-Dependent K+ Uptake by KtrB across the Plasma Membrane in Synechocystis sp. Strain PCC 6803 Peer-reviewed

    Lalu Zulkifli, Masaro Akai, Asuka Yoshikawa, Mie Shimojima, Hiroyuki Ohta, H. Robert Guy, Nobuyuki Uozumi

    JOURNAL OF BACTERIOLOGY　192　(19)　5063-5070　2010/10

    DOI： 10.1128/JB.00569-10 　

    ISSN： 0021-9193

  86. Roles of tandem-pore K plus channels in plants - a puzzle still to be solved Peer-reviewed

    C. Voelker, J. L. Gomez-Porras, D. Becker, S. Hamamoto, N. Uozumi, F. Gambale, B. Mueller-Roeber, K. Czempinski, I. Dreyer

    PLANT BIOLOGY　12　56-63　2010/09

    DOI： 10.1111/j.1438-8677.2010.00353.x 　

    ISSN： 1435-8603

  87. Cell-free protein N-myristoylation system utilizing wheat-germ extracts facilitates update of consensus motif for the modification of plant proteins Peer-reviewed

    Yamauchi, S, Fusada, N, Hayashi, H, Utsumi, T, Uozumi, N, Tozawa, Y

    FEBS. J.　2010/07/01

  88. Modulation of the Arabidopsis KAT1 channel by an activator of protein kinase C in Xenopus laevis oocytes Peer-reviewed

    Aiko Sato, Franco Gambale, Ingo Dreyer, Nobuyuki Uozumi

    FEBS JOURNAL　277　(10)　2318-2328　2010/05

    DOI： 10.1111/j.1742-4658.2010.07647.x 　

    ISSN： 1742-464X

    eISSN： 1742-4658

  89. Ion channels and plant stress: past, present and future Invited Peer-reviewed

    Uozumi, N, Schroeder J.I

    Ion Channels and Plant Stress Responses　1-22　2010/05/01

  90. A Trk/HKT-Type K+ Transporter from Trypanosoma brucei Peer-reviewed

    Marc Mosimann, Shinobu Goshima, Tanja Wenzler, Alexandra Luescher, Nobuyuki Uozumi, Pascal Maeser

    EUKARYOTIC CELL　9　(4)　539-546　2010/04

    DOI： 10.1128/EC.00314-09 　

    ISSN： 1535-9778

    eISSN： 1535-9786

  91. A Novel Potassium Channel in Photosynthetic Cyanobacteria Peer-reviewed

    Manuela Zanetti, Enrico Teardo, Nicoletta La Rocca, Lalu Zulkifli, Vanessa Checchetto, Toshiaki Shijuku, Yuki Sato, Giorgio Mario Giacometti, Noboyuki Uozumi, Elisabetta Bergantino, Ildiko Szabo

    PLOS ONE　5　(4)　1-10　2010/04

    DOI： 10.1371/journal.pone.0010118 　

    ISSN： 1932-6203

  92. Threonine at position 306 of the KAT1 potassium channel is essential for channel activity and is a target site for ABA-activated SnRK2/OST1/SnRK2.6 protein kinase Peer-reviewed

    Aiko Sato, Yuki Sato, Yoichiro Fukao, Masayuki Fujiwara, Taishi Umezawa, Kazuo Shinozaki, Takao Hibi, Mitsutaka Taniguchi, Hiroshi Miyake, Derek B. Goto, Nobuyuki Uozumi

    BIOCHEMICAL JOURNAL　424　439-448　2009/12

    DOI： 10.1042/BJ20091221 　

    ISSN： 0264-6021

    eISSN： 1470-8728

  93. The Implication of YggT of Escherichia coli in Osmotic Regulation Peer-reviewed

    Tomokazu Ito, Nobuyuki Uozumi, Tatsunosuke Nakamura, Sayuri Takayama, Nobuyuki Matsuda, Hirofumi Aiba, Hisashi Hemmi, Tohru Yoshimura

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY　73　(12)　2698-2704　2009/12

    DOI： 10.1271/bbb.90558 　

    ISSN： 0916-8451

    eISSN： 1347-6947

  94. Identification and Characterization of the Na+/H+ Antiporter Nhas3 from the Thylakoid Membrane of Synechocystis sp PCC 6803 Peer-reviewed

    Kenta Tsunekawa, Toshiaki Shijuku, Mitsuo Hayashimoto, Yoichi Kojima, Kiyoshi Onai, Megumi Morishita, Masahiro Ishiura, Teruo Kuroda, Tatsunosuke Nakamura, Hiroshi Kobayashi, Mayuko Sato, Kiminori Toyooka, Ken Matsuoka, Tatsuo Omata, Nobuyuki Uozumi

    JOURNAL OF BIOLOGICAL CHEMISTRY　284　(24)　16513-16521　2009/06

    DOI： 10.1074/jbc.M109.001875 　

    ISSN： 0021-9258

  95. Synchrony between flower opening and petal-color change from red to blue in morning glory, Ipomoea tricolor cv. Heavenly Blue Peer-reviewed

    Kumi Yoshida, Naoko Miki, Kazumi Mononoi, Miki Kawachi, Kiyoshi Katou, Yoshiji Okazaki, Nobuyuki Uozumi, Masayoshi Maeshima, Tadao Kondo

    PROCEEDINGS OF THE JAPAN ACADEMY SERIES B-PHYSICAL AND BIOLOGICAL SCIENCES　85　(6)　187-197　2009/06

    DOI： 10.2183/pjab.85.187 　

    ISSN： 0386-2208

  96. Contribution of salicylic acid glucosyltransferase, OsSGT1, to chemically induced disease resistance in rice plants Peer-reviewed

    Kenji Umemura, Junji Satou, Michiaki Iwata, Nobuyuki Uozumi, Jinichiro Koga, Tomonori Kawano, Tomokazu Koshiba, Hiroyuki Anzai, Masaaki Mitomi

    PLANT JOURNAL　57　(3)　463-472　2009/02

    DOI： 10.1111/j.1365-313X.2008.03697.x 　

    ISSN： 0960-7412

  97. Novel Treatment for Lithium-Induced Nephrogenic Diabetes Insipidus Rat Model Using the Sendai-Virus Vector Carrying Aquaporin 2 Gene Peer-reviewed

    Hidetaka Suga, Hiroshi Nagasaki, Taka-aki Kondo, Yoshiki Okajima, Chizuko Suzuki, Nobuaki Ozaki, Hiroshi Arima, Tokunori Yamamoto, Noriyuki Ozaki, Masaro Akai, Aiko Sato, Nobuyuki Uozumi, Makoto Inoue, Mamoru Hasegawa, Yutaka Oiso

    ENDOCRINOLOGY　149　(11)　5803-5810　2008/11

    DOI： 10.1210/en.2007-1806 　

    ISSN： 0013-7227

  98. Electrophysiological Properties of NtTPK1 Expressed in Yeast Tonoplast Peer-reviewed

    Shin Hamamoto, Isamu Yabe, Nobuyuki Uozumi

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY　72　(10)　2785-2787　2008/10

    DOI： 10.1271/bbb.80354 　

    ISSN： 0916-8451

    eISSN： 1347-6947

  99. Electrophysiological Properties of NtTPK1 Expressed in Yeast Tonoplast

    Shin Hamamoto, Isamu Yabe, Nobuyuki Uozumi

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY　72　(10)　2785-2787　2008/10

    DOI： 10.1271/bbb.80354 　

    ISSN： 0916-8451

    eISSN： 1347-6947

  100. Purification of the functional plant membrane channel KAT1 Peer-reviewed

    Takao Hibi, Shiho Aoki, Keisuke Oda, Shintaro Munemasa, Shunsuke Ozaki, Osamu Shirai, Yoshiyuki Murata, Nobuyuki Uozumi

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS　374　(3)　465-469　2008/09

    DOI： 10.1016/j.bbrc.2008.07.026 　

    ISSN： 0006-291X

    eISSN： 1090-2104

  101. Micropatterning of hydrogel and on-chip long time monitoring of individual cells in a cage

    Fumihito Arai, Hideyuki Matsumoto, Toshiaki Shijuku, Nobuyuki Uozumi

    12th International Conference on Miniaturized Systems for Chemistry and Life Sciences - The Proceedings of MicroTAS 2008 Conference　1861-1863　2008

    Publisher: Chemical and Biological Microsystems Society

  102. Characterization of a tobacco TPK-type K+ channel as a novel tonoplast K+ channel using yeast tonoplasts Peer-reviewed

    Shin Hamamoto, Junichiro Marui, Ken Matsuoka, Kyohei Higashi, Kazuei Igarashi, Tsuyoshi Nakagawa, Teruo Kuroda, Yasuo Mori, Yoshiyuki Murata, Yoichi Nakanishi, Masayoshi Maeshima, Isamu Yabe, Nobuyuki Uozumi

    JOURNAL OF BIOLOGICAL CHEMISTRY　283　(4)　1911-1920　2008/01

    DOI： 10.1074/jbc.M708213200 　

    ISSN： 0021-9258

    eISSN： 1083-351X

  103. Contribution of hydrophobic and electrostatic interactions to the membrane integration of the Shaker K+ channel voltage sensor domain Peer-reviewed

    Liyan Zhang, Yoko Sato, Tara Hessa, Gunnar von Heijne, Jong-Kook Lee, Itsuo Kodama, Masao Sakaguchi, Nobuyuki Uozumi

    PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA　104　(20)　8263-8268　2007/05

    DOI： 10.1073/pnas.0611007104 　

    ISSN： 0027-8424

  104. Role of positively charged amino acids in the M2(D) transmembrane helix of Ktr/Trk/HKT type cation transporters Peer-reviewed

    Naoki Kato, Masaro Akai, Lalu Zulkifli, Nobuyuki Matsuda, Yasuhiro Kato, Shinobu Goshima, Akihiro Hazama, Mutsumi Yamagami, H. Robert Guy, Nobuyuki Uozumi

    CHANNELS　1　(3)　161-171　2007/05

    ISSN： 1933-6950

  105. Mutation of his-157 in the second pore loop drastically reduces the activity of the Synechocystis Ktr-type transporter Peer-reviewed

    Lalu Zulkifli, Nobuyuki Uozumi

    JOURNAL OF BACTERIOLOGY　188　(22)　7985-7987　2006/11

    DOI： 10.1128/JB.00886-06 　

    ISSN： 0021-9193

  106. Nomenclature for HKT transporters, key determinants of plant salinity tolerance Peer-reviewed

    J. Damien Platten, Olivier Cotsaftis, Pierre Berthomieu, Hans Bohnert, Romola J. Davenport, David J. Fairbairn, Tomoaki Horie, Roger A. Leigh, Hong-Xuan Lin, Sheng Luan, Pascal Maeser, Omar Pantoja, Alonso Rodriguez-Navarro, Daniel P. Schachtman, Julian I. Schroeder, Herve Sentenac, Nobuyuki Uozumi, Anne-Alienor Very, Jian-Kang Zhu, Elizabeth S. Dennis, Mark Tester

    TRENDS IN PLANT SCIENCE　11　(8)　372-374　2006/08

    DOI： 10.1016/j.tplants.2006.06.001 　

    ISSN： 1360-1385

  107. Properties of Shaker-type potassium channels in higher plants Peer-reviewed

    F. Gambale, N. Uozumi

    JOURNAL OF MEMBRANE BIOLOGY　210　(1)　1-19　2006/03

    DOI： 10.1007/S00232-006-0856-X 　

    ISSN： 0022-2631

    eISSN： 1432-1424

  108. Ktr-mediated potassium transport, a major pathway for potassium uptake, is coupled to a proton gradient across the membrane in Synechocystis sp. PCC 6803 Peer-reviewed

    Nobuyuki Matsuda, Nobuyuki Uozumi

    Bioscience, Biotechnology and Biochemistry　70　(1)　273-275　2006

    DOI： 10.1271/bbb.70.273 　

    ISSN： 0916-8451 1347-6947

  109. All four putative selectivity filter glycine residues in KtrB are essential for high affinity and selective K+ uptake by the KtrAB system from Vibrio alginolyticus Peer-reviewed

    N Tholema, M Vor der Bruggen, P Maser, T Nakamura, JI Schroeder, H Kobayashi, N Uozumi, EP Bakker

    JOURNAL OF BIOLOGICAL CHEMISTRY　280　(50)　41146-41154　2005/12

    DOI： 10.1074/jbc.M507647200 　

    ISSN： 0021-9258

  110. Enhanced salt tolerance mediated by AtHKT1 transporter-induced Na+ unloading from xylem vessels to xylem parenchyma cells Peer-reviewed

    Sunarpi, T Horie, J Motoda, M Kubo, H Yang, K Yoda, R Horie, WY Chan, HY Leung, K Hattori, M Konomi, M Osumi, M Yamagami, JI Schroeder, N Uozumi

    PLANT JOURNAL　44　(6)　928-938　2005/12

    DOI： 10.1111/j.1365-313X.2005.02595.x 　

    ISSN： 0960-7412

  111. Further application of a two-step heparin affinity chromatography method using divalent cations as eluents: Purification and identification of membrane-bound heparin binding proteins from the mitochondrial fraction of HL-60 cells Peer-reviewed

    T Iida, M Kamo, N Uozumi, T Inui, K Imai

    JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES　823　(2)　209-212　2005/09

    DOI： 10.1016/j.chroma.2005.06.025 　

    ISSN： 1570-0232

  112. Na+-dependent K+ uptake ktr system from the cyanobacterium Synechocystis sp PCC 6803 and its role in the early phases of cell adaptation to hyperosmotic shock Peer-reviewed

    N Matsuda, H Kobayashi, H Katoh, T Ogawa, L Futatsugi, T Nakamura, EP Bakker, N Uozumi

    JOURNAL OF BIOLOGICAL CHEMISTRY　279　(52)　54952-54962　2004/12

    DOI： 10.1074/jbc.M407268200 　

    ISSN： 0021-9258

  113. Mechanosensitivity of GIRK channels is mediated by protein kinase C-dependent channel-phosphatidylinositol 4,5-bisphosphate interaction Peer-reviewed

    LY Zhang, JK Lee, SA John, N Uozumi, Kodama, I

    JOURNAL OF BIOLOGICAL CHEMISTRY　279　(8)　7037-7047　2004/02

    DOI： 10.1074/jbc.M307323200 　

    ISSN： 0021-9258

  114. Topogenesis of two transmembrane type K+ channels, Kir 2.1 and KcsA Peer-reviewed

    N Umigai, Y Sato, A Mizutani, T Utsumi, M Sakaguchi, N Uozumi

    JOURNAL OF BIOLOGICAL CHEMISTRY　278　(41)　40373-40384　2003/10

    DOI： 10.1074/jbc.M307451200 　

    ISSN： 0021-9258

  115. Addition of a peptide tag at the C terminus of AtHKT1 inhibits its Na+ transport Peer-reviewed

    Y Kato, A Hazama, M Yamagami, N Uozumi

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY　67　(10)　2291-2293　2003/10

    DOI： 10.1271/bbb.67.2291 　

    ISSN： 0916-8451

    eISSN： 1347-6947

  116. Functional analysis of AtHKT1 in Arabidopsis shows that Na+ recirculation by the phloem is crucial for salt tolerance Peer-reviewed

    Pierre Berthomieu, Geneviève Conéjéro, Aurélie Nublat, William J. Brackenbury, Cécile Lambert, Cristina Savio, Nobuyuki Uozumi, Shigetoshi Oiki, Katsuyuki Yamada, Françoise Cellier, Françoise Gosti, Thierry Simonneau, Pauline A. Essah, Mark Tester, Anne-Aliénor Véry, Hervé Sentenac, Francine Casse

    EMBO Journal　22　(9)　2004-2014　2003/05/01

    DOI： 10.1093/emboj/cdg207 　

    ISSN： 0261-4189

  117. Molecular dissection of the contribution of negatively and positively charged residues in S2, S3, and S4 to the final membrane topology of the voltage sensor in the K+ channel, KAT1 Peer-reviewed

    Y Sato, M Sakaguchi, S Goshima, T Nakamura, N Uozumi

    JOURNAL OF BIOLOGICAL CHEMISTRY　278　(15)　13227-13234　2003/04

    DOI： 10.1074/jbc.M300431200 　

    ISSN： 0021-9258

  118. KtrAB and KtrCD: Two K+ uptake systems in Bacillus subtilis and their role in adaptation to hypertonicity Peer-reviewed

    G Holtmann, EP Bakker, N Uozumi, E Bremer

    JOURNAL OF BACTERIOLOGY　185　(4)　1289-1298　2003/02

    DOI： 10.1128/JB.185.4.1289-1298.2003 　

    ISSN： 0021-9193

  119. Requirement of Negative Residues, Asp 95 and Asp 105, in S2 on Membrane Integration of a Voltage-dependent K+Channel, KAT1 Peer-reviewed

    Yoko SATO, Yoshihiro HOSOO, Masao SAKAGUCHI, Nobuyuki UOZUMI

    Bioscience, Biotechnology, and Biochemistry　67　(4)　923-926　2003/01

    Publisher: Informa UK Limited

    DOI： 10.1271/bbb.67.923 　

    ISSN： 0916-8451

    eISSN： 1347-6947

  120. Membrane-bound heparin binding proteins from HL-60 cells purified in a two-step affinity chromatography differentially eluted with divalent cations Peer-reviewed

    Katsuyuki Imai, Tsukimi Iida, Yasuo Takano, Nobuyuki Uozumi

    Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences　780　(1)　1-12　2002/11/15

    DOI： 10.1016/S1570-0232(02)00404-X 　

    ISSN： 1570-0232

  121. Altered shoot/root Na+ distribution and bifurcating salt sensitivity in Arabidopsis by genetic disruption of the Na+ transporter AtHKT1 Peer-reviewed

    Pascal Mäser, Brendan Eckelman, Rama Vaidyanathan, Tomoaki Horie, David J Fairbairn, Masahiro Kubo, Mutsumi Yamagami, Katsushi Yamaguchi, Mikio Nishimura, Nobuyuki Uozumi, Whitney Robertson, Michael R Sussman, Julian I Schroeder

    FEBS Letters　531　(2)　157-161　2002/11/06

    DOI： 10.1016/S0014-5793(02)03488-9 　

    ISSN： 0014-5793

  122. Glycine residues in potassium channel-like selectivity filters determine potassium selectivity in four-loop-per-subunit HKT transporters from plants Peer-reviewed

    P Maser, Y Hosoo, S Goshima, T Horie, B Eckelman, K Yamada, K Yoshida, EP Bakker, A Shinmyo, S Oiki, JI Schroeder, N Uozumi

    PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA　99　(9)　6428-6433　2002/04

    DOI： 10.1073/pnas.082123799 　

    ISSN： 0027-8424

  123. Integration of Shaker-type K+ channel, KAT1, into the endoplasmic reticulum membrane: Synergistic insertion of voltage-sensing segments, S3-S4, and independent insertion of pore-forming segments, S5-P-S6 Peer-reviewed

    Y Sato, M Sakaguchi, S Goshima, T Nakamura, N Uozumi

    PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA　99　(1)　60-65　2002/01

    DOI： 10.1073/pnas.012399799 　

    ISSN： 0027-8424

  124. Residue aspartate-147 from the third transmembrane region of Na+/H+ antiporter NhaB of Vibrio alginolyticus plays a role in its activity Peer-reviewed

    T Nakamura, Y Fujisaki, H Enomoto, Y Nakayama, T Takabe, N Yamaguchi, N Uozumi

    JOURNAL OF BACTERIOLOGY　183　(19)　5762-5767　2001/10

    DOI： 10.1128/JB.183.19.5762-5767.2001 　

    ISSN： 0021-9193

  125. Sodium blocking induced by a point mutation at the C-terminal end of the pore helix of KAT1 channel. Peer-reviewed

    Uozumi, N, Yamada, K, Goshima, S, Ona, T, Oiki, S

    J. Membrane Biol.　181,　163-170　2001/10

  126. Molecular mechanisms of potassium and sodium transport in plants Peer-reviewed

    Schroeder, J, Buschmann, P, Eckelman, B, Kim, E, Sussman, M, Uozumi, N, Maser, P

    Plant Nutrition　92　2001/06/01

    DOI： 10.1023/A:1021159130729 　

  127. Evidence in support of a four transmembranepore-transmembrane topology model for the Arabidopsis thaliana Na+/K+ translocating AtHKT1 protein, a member of the superfamily of k+ transporters Peer-reviewed

    Yasuhiro Kato, Masao Sakaguchi, Yasuo Mori, Kumiko Saito, Tatsunosuke Nakamura, Evert P. Bakker, Yoko Sato, Shinobu Goshima, Nobuyuki Uozumi

    Proceedings of the National Academy of Sciences of the United States of America　98　(11)　6488-6493　2001/05/22

    DOI： 10.1073/pnas.101556598 　

    ISSN： 0027-8424

  128. Escherichia coli as an expression system for K+ transport systems from plants Peer-reviewed

    N. Uozumi

    American Journal of Physiology - Cell Physiology　281　(3)　C733-C739　2001

    ISSN： 0363-6143

  129. Involvement of apoplastic peroxidases in the chitosaccharide-induced immediate oxidative bursts and a cytosolic Ca2+ increase in tobacco suspension culture. Peer-reviewed

    Kawano, T, Sahashi, N, Uozumi, N, Muto, S

    Plant Peroxidase Newsletter　14　14-14　2000/10

  130. The Arabidopsis HKT1 gene homologue mediates inward Na+ currents in Xenopus oocytes and Na+ uptake in Saccharomyces cerevisiae. Peer-reviewed

    Uozumi, N, Kim, E.J, Rubio, F, Yamaguchi, T, Muto, S, Tsuboi, A, Bakker, E.P, Nakamura, T, Schroeder, J.I

    Plant Physiol.　122　(4)　1249-1259,　2000/10

  131. Cloning of a cDNA encoding a 66-kDa Ca2+-dependent protein kinase (CDPK) from Dunaliella tertiolecta (Chlorophyta) Peer-reviewed

    Reinhard Pinontoan, Takashi Yuasa, Marinela I. Anderca, Takashi Matsuoka, Nobuyuki Uozumi, Hitoshi Mori, Shoshi Muto

    Journal of Phycology　36　(3)　545-552　2000/06

    DOI： 10.1046/j.1529-8817.2000.99185.x 　

    ISSN： 0022-3646

  132. yam8+, a Schizosaccharomyces pombe gene, is a potential homologue of the Saccharomyces cerevisiae MID1 gene encoding a stretch-activated Ca2+-permeable channel Peer-reviewed

    Yasushi Tasaka, Yuko Nakagawa, Chikara Sato, Masanobu Mino, Nobuyuki Uozumi, Norio Murata, Shoshi Muto, Hidetoshi Iida

    Biochemical and Biophysical Research Communications　269　(1)　265-269　2000/03/05

    Publisher: Academic Press Inc.

    DOI： 10.1006/bbrc.2000.2278 　

    ISSN： 0006-291X

  133. Aromatic monoamine-induced immediate oxidative burst leading to an increase in cytosolic Ca2+ concentration in tobacco suspension culture Peer-reviewed

    Tomonori Kawano, Reinhard Pinontoan, Nobuyuki Uozumi, Chikahiro Miyake, Kozi Asada, Pappachan E. Kolattukudy, Shoshi Muto

    Plant and Cell Physiology　41　(11)　1251-1258　2000

    Publisher: Japanese Society of Plant Physiologists

    DOI： 10.1093/pcp/pcd052 　

    ISSN： 0032-0781

  134. Phenylethylamine-induced generation of reactive oxygen species and ascorbate free radicals in tobacco suspension culture: Mechanism for oxidative burst mediating Ca2+ influx Peer-reviewed

    Tomonori Kawano, Reinhard Pinontoan, Nobuyuki Uozumi, Yasujiro Morimitsu, Chikahiro Miyake, Kozi Asada, Shoshi Muto

    Plant and Cell Physiology　41　(11)　1259-1266　2000

    Publisher: Japanese Society of Plant Physiologists

    DOI： 10.1093/pcp/pcd053 　

    ISSN： 0032-0781

  135. Phosphorylation of the inward rectifying potassium channel KAT1 by ABR kinase in Vicia guard cells Peer-reviewed

    Izumi C. Mori, Nobuyuki Uozumi, Shoshi Muto

    Plant and Cell Physiology　41　(7)　850-856　2000

    Publisher: Japanese Society of Plant Physiologists

    DOI： 10.1093/pcp/pcd003 　

    ISSN： 0032-0781

  136. Crystallization and preliminary X-ray analysis of β-amylase from Bacillus polymyxa Peer-reviewed

    Takashi Yamane, Hiroshi Tasaki, Fusako Matsumoto, Atsuo Suzuki, Nobuyuki Uozumi, Tamaichi Ashida

    Acta Crystallographica Section D: Biological Crystallography　55　(4)　898-900　1999/04

    DOI： 10.1107/S0907444998017570 　

    ISSN： 0907-4449

  137. Suppression of inward-rectifying K+ channels KAT1 and AKT2 by dominant negative point mutations in the KAT1 α-subunit Peer-reviewed

    V. M. Baizabal-Aguirre, S. Clemens, N. Uozumi, J. I. Schroeder

    Journal of Membrane Biology　167　(2)　119-125　1999

    DOI： 10.1007/s002329900476 　

    ISSN： 0022-2631

  138. Development of rotating-mesh basket type bioreactor for carrot embryo production in immobilized callus system. Peer-reviewed

    Suehara, K, Nagamori, E, Honda, H, Uozumi, N, Kobayashi, T

    J. Chem. Eng. Japan,　31　(4)　613-617.　1998/10

    DOI： 10.1252/jcej.31.613 　

  139. Salicylic acid induces a cytosolic Ca2+ elevation in yeast. Peer-reviewed

    Mori, I. C, Iida, H, Tsuji, F. I, Isobe, M, Uozumi, N, Muto, S

    Biosci. Biotech. Biochem.　62　(5)　986-989.　1998/10

    Publisher: Japan Society for Bioscience, Biotechnology, and Agrochemistry

    DOI： 10.1271/bbb.62.986 　

    ISSN： 0916-8451

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    Cytosolic free calcium ion concentration ([Ca^<2+>]_<cyt>) after a salicylic acid (SA)-stimulus was monitored in cells of the yeast Saccharomyces cerevisiae expressing apoaequorin, which constitutes a Ca^<2+>-sensitive luminescent protein, aequorin, when combined with coelenterazine. SA induced a transient [Ca^<2+>]_<cyt> elevation that was dependent on the concentration of SA and pH of the SA solution. The SA-induced [Ca^<2+>]_<cyt> elevation was not reduced in Ca^<2+>-deficient medium, suggesting that Ca^<2+> was mobilized from an intracellular Ca^<2+> store (s). Benzoic acid, butyric acid and sorbic acid did not induced a [Ca^<2+>]_<cyt> elevation.

  140. Determination of transmembrane topology of an inward-rectifying potassium channel from Arabidopsis thaliana based on functional expression in Escherichia coli Peer-reviewed

    Nobuyuki Uozumi, Tatsunosuke Nakamura, Julian I. Schroeder, Shoshi Muto

    Proceedings of the National Academy of Sciences of the United States of America　95　(17)　9773-9778　1998/08/18

    DOI： 10.1073/pnas.95.17.9773 　

    ISSN： 0027-8424

  141. Salicylic acid induces extracellular superoxide generation followed by an increase in cytosolic calcium ion in tobacco suspension culture: The earliest events in salicylic acid signal transduction Peer-reviewed

    Tomonori Kawano, Nobuya Sahashi, Koji Takahashi, Nobuyuki Uozumi, Shoshi Muto

    Plant and Cell Physiology　39　(7)　721-730　1998

    Publisher: Oxford University Press

    DOI： 10.1093/oxfordjournals.pcp.a029426 　

    ISSN： 0032-0781

  142. AtKUP1: An arabidopsis gene encoding high-affinity potassium transport activity Peer-reviewed

    Eugene J. Kim, June Myoung Kwak, Nobuyuki Uozumi, Julian I. Schroeder

    Plant Cell　10　(1)　51-62　1998

    Publisher: American Society of Plant Biologists

    DOI： 10.1105/tpc.10.1.51 　

    ISSN： 1040-4651

  143. Production of regenerated plantlet using shaking vessel-type bioreactor Peer-reviewed

    Hiroyuki Honda, Toshiyuki Hattori, Nobuyuki Uozumi, Takeshi Kobayashi, Yoshihito Kato, Setsuro Hiraoka

    Journal of Chemical Engineering of Japan　30　(1)　179-182　1997

    Publisher: Society of Chemical Engineers, Japan

    DOI： 10.1252/jcej.30.179 　

    ISSN： 0021-9592

  144. Plant regeneration and somatic embryogenesis frequency using callus induced from regenerated celery plant Peer-reviewed

    K Suehara, S Goto, N Uozumi, T Kobayashi

    KAGAKU KOGAKU RONBUNSHU　22　(3)　691-694　1996/05

    ISSN： 0386-216X

  145. Optimal expression of GUS gene from methyl jasmonate-inducible promoter in high density culture of transformed tobacco cell line BY-2 Peer-reviewed

    KI Suehara, S Takao, K Nakamura, N Uozumi, T Kobayashi

    JOURNAL OF FERMENTATION AND BIOENGINEERING　82　(1)　51-55　1996

    DOI： 10.1016/0922-338X(96)89454-2 　

    ISSN： 0922-338X

  146. Efficient regeneration from GUS-transformed Ajuga hairy root Peer-reviewed

    N Uozumi, Y Ohtake, Y Nakashimada, Y Morikawa, N Tanaka, T Kobayashi

    JOURNAL OF FERMENTATION AND BIOENGINEERING　81　(5)　374-378　1996

    DOI： 10.1016/0922-338X(96)85135-X 　

    ISSN： 0922-338X

  147. Efficient culture method for production of plantlets from mechanically cut horseradish hairy roots Peer-reviewed

    Y Nakashimada, N Uozumi, T Kobayashi

    JOURNAL OF FERMENTATION AND BIOENGINEERING　81　(1)　87-89　1996

    DOI： 10.1016/0922-338X(96)83128-X 　

    ISSN： 0922-338X

  148. Multuple genes, tissue specificity, and expression-dependent modulation contribute to the functional diversity of potassium channels in Arabidopsis thaliana. Peer-reviewed

    Cao. Y, Ward, J. M, Kelly, W. B, Ichida, A. M, Gaber, R. F, Anderson, J. A, Uozumi, N, Schroeder, J. I, Crawford. N. M

    Plant Physiol.　109,　1093-1106.　1995/10

    DOI： 10.1104/pp.109.3.1093 　

  149. IDENTIFICATION OF STRONG MODIFICATIONS IN CATION SELECTIVITY IN AN ARABIDOPSIS INWARD RECTIFYING POTASSIUM CHANNEL BY MUTANT SELECTION IN YEAST Peer-reviewed

    N UOZUMI, W GASSMANN, YW GAO, JI SCHROEDER

    JOURNAL OF BIOLOGICAL CHEMISTRY　270　(41)　24276-24281　1995/10

    DOI： 10.1074/jbc.270.41.24276 　

    ISSN： 0021-9258

  150. 20-HYDROXYECDYSONE PRODUCTION IN AJUGA HAIRY ROOT CONTROLLING INTRACELLULAR PHOSPHATE CONTENT-BASED ON KINETIC-MODEL Peer-reviewed

    N UOZUMI, S MAKINO, T KOBAYASHI

    JOURNAL OF FERMENTATION AND BIOENGINEERING　80　(4)　362-368　1995

    DOI： 10.1016/0922-338X(95)94205-6 　

    ISSN： 0922-338X

  151. EFFICIENT PRODUCTION OF CELERY EMBRYOS AND PLANTLETS RELEASED IN CULTURE OF IMMOBILIZED GEL BEADS Peer-reviewed

    KI SUEHARA, K KOHKETSU, N UOZUMI, T KOBAYASHI

    JOURNAL OF FERMENTATION AND BIOENGINEERING　79　(6)　585-588　1995

    DOI： 10.1016/0922-338X(95)94752-D 　

    ISSN： 0922-338X

  152. PRODUCTION OF PLANTLETS FOR USE AS ARTIFICIAL SEEDS FROM HORSERADISH HAIRY ROOTS FRAGMENTED IN A BLENDER Peer-reviewed

    Y NAKASHIMADA, N UOZUMI, T KOBAYASHI

    JOURNAL OF FERMENTATION AND BIOENGINEERING　79　(5)　458-464　1995

    DOI： 10.1016/0922-338X(95)91262-4 　

    ISSN： 0922-338X

  153. Micropropagation of horseradish hairy root by means of adventitious shoot primordia. Peer-reviewed

    Uozumi, N, Asano, Y, Kobayashi, T

    Plant Cell Tissue Organ Culture　36　183-190.　1994/10

  154. Light activation of expression associated with the tomato rbcS promoter in transformed tobacco cell line BY-2 Peer-reviewed

    Nobuyuki Uozumi, Yoshihisa Inoue, Ken-ichi Yamazaki, Takeshi Kobayashi

    Journal of Biotechnology　36　(1)　55-62　1994/07/29

    DOI： 10.1016/0168-1656(94)90023-X 　

    ISSN： 0168-1656

  155. オーキシンによる成長点形成促進効果を利用した毛状根の効率的培養および増殖モデルの構築. Invited

    中島田豊, 魚住信之, 小林 猛

    化学工学シンポジウムシリーズ　37　82-87　1994

  156. MOLECULAR-CLONING OF THERMOSTABLE BETA-GLUCOSIDASE GENE FROM A THERMOPHILIC ANAEROBE NA10 AND ITS HIGH EXPRESSION IN ESCHERICHIA-COLI Peer-reviewed

    H SOTA, P ARUNWANICH, O KURITA, N UOZUMI, H HONDA, S IIJIMA, T KOBAYASHI

    JOURNAL OF FERMENTATION AND BIOENGINEERING　77　(2)　199-201　1994

    DOI： 10.1016/0922-338X(94)90324-7 　

    ISSN： 0922-338X

  157. STIMULATION OF EMERGENCE OF ROOT APICAL MERISTEMS IN HORSERADISH HAIRY ROOT BY AUXIN SUPPLEMENTATION AND ITS KINETIC-MODEL Peer-reviewed

    Y NAKASHIMADA, N UOZUMI, T KOBAYASHI

    JOURNAL OF FERMENTATION AND BIOENGINEERING　77　(2)　178-182　1994

    DOI： 10.1016/0922-338X(94)90320-4 　

    ISSN： 0922-338X

  158. Light dependency in celery somatic embryogenesis and plantlet development in suspension culture. Peer-reviewed

    Uozumi, N, Kohketsu, K, Okamoto, A, Kobayashi, T

    Plant Tissue Culture Lett.　10　(1)　25-32.　1993/10

    Publisher: Japanese Society for Plant Cell and Molecular Biology

    DOI： 10.5511/plantbiotechnology1984.10.25 　

    ISSN： 0289-5773

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    We analyzed the light requirements for celery somatic embryogenesis and plantlet development. The regeneration period was divided into two stages. During the first stage, in which torpedo-shaped embryos were mainly produced from the embryogenic callus, light stimulated chlorophyll synthesis. Although under dark conditions cells were slow in embryo formation, prolonged culture in the dark led to the production of globular-, heart- and torpedo-shaped embryos. The torpedo embryos were ready to become healthy plantlets after the second-stage culture under light conditions. Light enhanced the embryo formation rate, but was not obligatory on embryo formation. In the second stage, a 16-h photoperiod and a higher light intensity gave the maximum frequency of Plantlet development, and were essential for the production of the healthy plantlets in terms of size (more than 850μm). The results strongly suggest that illumination can be reduced at the beginning of the first regeneration stage, but that increased illumination is required after the torpedo embryo development has progressed.

  159. APPLICATION OF IMAGE-ANALYSIS WITH NEURAL-NETWORK FOR PLANT SOMATIC EMBRYO CULTURE Peer-reviewed

    N UOZUMI, T YOSHINO, S SHIOTANI, K SUEHARA, F ARAI, T FUKUDA, T KOBAYASHI

    JOURNAL OF FERMENTATION AND BIOENGINEERING　76　(6)　505-509　1993

    DOI： 10.1016/0922-338X(93)90249-8 　

    ISSN： 0922-338X

  160. SECRETION OF THERMOPHILIC BACTERIAL CELLOBIOHYDROLASE IN SACCHAROMYCES-CEREVISIAE Peer-reviewed

    N UOZUMI, A HAYASHI, T ITO, A PATTHRA, YAMASHITA, I, S IIJIMA, T KOBAYASHI

    JOURNAL OF FERMENTATION AND BIOENGINEERING　75　(6)　399-404　1993

    DOI： 10.1016/0922-338X(93)90084-L 　

    ISSN： 0922-338X

  161. CLONING AND SEQUENCING OF A GENE ENCODING NITRITE REDUCTASE FROM PARACOCCUS-DENITRIFICANS AND EXPRESSION OF THE GENE IN ESCHERICHIA-COLI Peer-reviewed

    T OHSHIMA, M SUGIYAMA, N UOZUMI, S IIJIMA, T KOBAYASHI

    JOURNAL OF FERMENTATION AND BIOENGINEERING　76　(2)　82-88　1993

    DOI： 10.1016/0922-338X(93)90061-C 　

    ISSN： 0922-338X

  162. GROWTH AND KINETIC-PARAMETERS OF AJUGA HAIRY ROOT IN FED-BATCH CULTURE ON MONOSACCHARIDE MEDIUM Peer-reviewed

    N UOZUMI, K KOHKETSU, T KOBAYASHI

    JOURNAL OF CHEMICAL TECHNOLOGY AND BIOTECHNOLOGY　57　(2)　155-161　1993

    DOI： 10.1002/jctb.280570210 　

    ISSN： 0268-2575

  163. Excretion of peroxidase from horseradish hairy root in combination with ion supplementation. Peer-reviewed

    Uozumi, N, Kato, Y, Nakashimada, Y, Kobayashi, T

    Appl. Microbiol. Biotechnol　37　(5)　560-565.　1992/10

  164. Inducible production of recombinant xylose isomerase by escherichia coli in fed-batch culture Peer-reviewed

    Tomoyasu Kawabe, Takayuki Ohshima, Nobuyuki Uozumi, Shinji Iijima, Takeshi Kobayashi

    Journal of Chemical Engineering of Japan　25　(6)　702-708　1992

    DOI： 10.1252/jcej.25.702 　

    ISSN： 0021-9592

  165. PRODUCTION OF ARTIFICIAL SEED FROM HORSERADISH HAIRY ROOT Peer-reviewed

    N UOZUMI, Y NAKASHIMADA, Y KATO, T KOBAYASHI

    JOURNAL OF FERMENTATION AND BIOENGINEERING　74　(1)　21-26　1992

    DOI： 10.1016/0922-338X(92)90262-S 　

    ISSN： 0922-338X

  166. Enhancement of peroxidase production and excretion from horseradish hairy roots by light, NaCl and peroxidase-adsorption in situ. Peer-reviewed

    Kato, Y, Uozumi, N, Kimura, T, Honda, H, Kobayashi, T

    Plant Tissue Culture Lett.　8　(3)　158-165.　1991/10

    Publisher: Japanese Society for Plant Cell and Molecular Biology

    DOI： 10.5511/plantbiotechnology1984.8.158 　

    ISSN： 0289-5773

    More details Close

    Effects of light, NaCl and adsorption in situ on production and recovery of peroxidase from horseradish hairy root cell are reported in this paper. Illumination with fluorescent white light increased peroxidase content in the cell 2-fold. The treatment with NaCl stimulated excretion of peroxidase from the cell, and it was also subject to alteration in morphology without serious influence on proliferation. Adsorption of peroxidase from the extracellular medium with hydrophobic resin greatly enhanced the release of peroxidase. For a 43-day culture period, 70U/ml of peroxidase was produced compared to 5U/ml without special treatments. These results suggest that production and product recovery from plant cells can be greatly improved by application of the combined treatments.

  167. Structural and Functional Roles of Cysteine Residues of Bacillus polymyxa β-Amylase Peer-reviewed

    Nobuyuki Uozumi, Tsukasa Matsuda, Norihiro Tsukagoshi, Shigezo Udaka

    Biochemistry　30　(18)　4594-4599　1991/05/01

    DOI： 10.1021/bi00232a033 　

    ISSN： 1520-4995 0006-2960

  168. FED-BATCH CULTURE OF HAIRY ROOT USING FRUCTOSE AS A CARBON SOURCE Peer-reviewed

    N UOZUMI, K KOHKETSU, O KONDO, H HONDA, T KOBAYASHI

    JOURNAL OF FERMENTATION AND BIOENGINEERING　72　(6)　457-460　1991

    DOI： 10.1016/0922-338X(91)90054-K 　

    ISSN： 0922-338X

  169. Proteases involved in generation of β- and α-amylases from a large amylase precursor in Bacillus polymyxa Peer-reviewed

    S. Takekawa, N. Uozumi, N. Tsukagoshi, S. Udaka

    Journal of Bacteriology　173　(21)　6820-6825　1991

    DOI： 10.1128/jb.173.21.6820-6825.1991 　

    ISSN： 0021-9193

  170. A single gene directs synthesis of a precursor protein with b- and a-amylase activities in Bacillus polymyxa. Peer-reviewed

    Uozumi, N, Sakurai, K, Sasaki, T, Takekawa, S, Yamagata, H, Tsukagoshi, N, Udaka, S

    J. Bacteriol.　171　(1)　375-382.　1989/10

  171. Cloning and nucleotide sequence of the gene coding for enzymatically active fragments of the Bacillus polymyxa β-amylase Peer-reviewed

    T. Kawazu, Y. Nakanishi, N. Uozumi, T. Sasaki, H. Yamagata, N. Tsukagoshi, S. Udaka

    Journal of Bacteriology　169　(4)　1564-1570　1987

    ISSN： 0021-9193

Show all ︎Show first 5

Misc. 39

  1. AtHKT1 alleviates Na accumulation in companion cells of filaments during reproductive development in saline environments

    UCHIYAMA Takeshi, SAITO Shunya, YAMANASHI Taro, KATO Megumi, TAKAGI Tomoko, NAGATA Noriko, TOYAMA Sho, MIWA Misako, MATSUYAMA Shigeo, IKEDA Hayato, KIKUNAGA Hidetoshi, SUDA Toshimi, TSUJII Masaru, ISHIMARU Yasuhiro, UOZUMI Nobuyuki

    日本植物生理学会年会(Web)　65th　2024

  2. Analysis of K and Na transporters in plants

    内山剛志, 山梨太郎, 池田隼人, 菊永英寿, 石丸泰寛, 魚住信之

    アイソトープ・放射線研究発表会(Web)　60th　2023

    ISSN： 2436-4487

  3. Na輸送体AtHKT1の生殖時における耐塩性機構の解明

    内山剛志, 加藤恵, 池田隼人, 菊永英寿, 須田利美, 石丸泰寛, 魚住信之

    日本土壌肥料学会講演要旨集(Web)　69　2023

    ISSN： 2424-0575

  4. K+チャネル阻害剤による孔辺細胞アクチン繊維の構造変化

    上村明日香, 佐藤奏音, 石丸泰寛, 魚住信之, 檜垣匠

    日本植物学会大会研究発表記録(CD-ROM)　87th　2023

  5. Analysis of potassium ion transporters expressed in the shoot tissues

    山梨太郎, 東大起, 内山剛志, 白川由美子, 池田隼人, 菊永英寿, 須田利美, 山上睦, 辻井雅, 石丸泰寛, 魚住信之

    日本植物生理学会年会(Web)　63rd　2022

  6. Biosynthesis pathway of reactive persulfides in photosynthetic bacteria

    JUNG Minkyung, 雨宮大雅, 井田智章, 解良康太, 西村明, 本橋ほづみ, 魚住信之, 赤池孝章

    日本細菌学雑誌(Web)　74　(1)　2019

    ISSN： 1882-4110

  7. シアノバクテリアSynechocystis sp.PCC6803の塩ストレス誘導性バイオフィルム形成に必要なヒスチジンキナーゼの同定とシグナル伝達機構

    永山達也, 久米井智裕, 牧野恒平, 井田智章, 赤池孝章, 鈴木石根, 七谷圭, 魚住信之

    日本農芸化学会大会講演要旨集(Web)　2015　2015

    ISSN： 2186-7976

  8. ラン藻の塩誘導性バイオフィルム形成に関与する二成分系c-di-GMP合成酵素の同定

    牧野恒平, 七谷圭, 井田智章, 澤智裕, 澤智裕, 佐伯千香, 鈴木石根, 兵藤守, 早川芳弘, 赤池孝章, 魚住信之

    日本分子生物学会年会プログラム・要旨集(Web)　37th　2014

  9. 56. Jasmonate Glucoside: COI1-JAZs independent induction of leaf-closing movement in Samanea saman

    Ueda Minoru, Nakamura Yoko, Hamamoto Shin, Uozumi Nobuyuki, Boland Wilhelm, Mithofer Axel, Kombrink Erich

    (45)　73-73　2010/10/01

    Publisher: The Japanese Society for Chemical Regulation of Plants

    ISSN： 0919-1887

    More details Close

    We recently identified jasmonate glucoside (Leaf-closing Factor: LCF), a β-D-glucopyranoside of 12-OH JA, as a bioactive metabolite controlling nyctinastic leaf movement of Samanea saman, which has been used as a model plant in the studies on nyctinastic leaf movement of legumes. The bioactivity of LCF is completely different from other jasmonate derivatives. Here, we demonstrate that JA activity mediated through the COI1-JAZs signalling module and leaf-closing activity (with cell-shrinking acitivity) of JA glucoside might belong to independent signalling pathways. However, COI1-JAZ-dependent and independent signaling pathways may exists in the same plant side by side, as our results demonstrate because S. saman is responding to both LCF and JA in different mechanisms based on independent signalling modules. Our result implies the existence of an additional mode of action on jasmonates.

  10. The consensus motif for N-myristoylation of plant proteins with a wheat germ cell-free translation system

    YAMAUCHI Seiji, FUSADA Naoki, FUSADA Naoki, HAYASHI Hidenori, UTSUMI Toshihiko, UOZUMI Nobuyuki, ENDO Yaeta, ENDO Yaeta, TOZAWA Yuzuru

    FEBS Journal　277　(17)　3596-3607　2010

    DOI： 10.1111/j.1742-4658.2010.07768.x 　

    ISSN： 1742-464X

  11. Involvement of Aquaporin in Photomixotrophic Growth of Synechocystis sp. PCC 6803

    Akai Masaro, Onai Kiyoshi, Morishita Megumi, Kusano Miyako, Sato Mayuko, Redestig Henning, Kobayashi Makoto, Otsuki Hitomi, Matsuoka Ken, Toyooka Kiminori, Saito Kazuki, Ishiura Masahiro, Uozumi Nobuyuki

    Plant and Cell Physiology Supplement　2010　(0)　764-764　2010

    Publisher: The Japanese Society of Plant Physiologists

    More details Close

    Aquaporin is a water permeable channel, which has been found in bacteria, archaea and eukaryotic cells. The physiological role in cyanobacteria is not fully understood. &lt;I&gt;Synechocystis&lt;/I&gt; sp. PCC 6803 genome contains a single member of this gene family. We have previously evaluated the function of &lt;I&gt;Synechocystis&lt;/I&gt; aquaporin using &lt;I&gt;Xenopus&lt;/I&gt; oocytes expression system and &lt;I&gt;Synechocystis&lt;/I&gt; aquaporin deficient mutant on osmoregulation. In this study, we have studied the effect of glucose on the growth of the mutant. The photoautotrophic growth rates of the wild type and the mutant were identical. However, when grown photomixotrophically with glucose, the mutant ceased the growth within 48h. Furthermore, addition of glucose to the mutant resulted in elevated glycogen level, and triggered the structural aberrations and morphological abnormalities. These results demonstrate that aquaporin is essential for the growth of &lt;I&gt;Synechocystis&lt;/I&gt; sp. PCC 6803 under photomixotrophic conditions.

  12. Ion channels and plant stress: past, present and future

    Uozumi, N, Schroeder J.I

    Ion Channels and Plant Stress Responses　1-22　2010

    Publisher: Springer-Verlag Berlin Heidelberg

  13. Production of human secreted alkaline phosphatase in suspension and immobilization cultures of tobacco NT1 cells

    Naorni Shibasaki-Kitakawa, Takuya Miyamoto, Masaki Kubo, Nobuyuki Uozumi, Kinya Toriyama, Toshikuni Yonemoto, Michael L. Shuler

    JOURNAL OF BIOSCIENCE AND BIOENGINEERING　108　S5-S5　2009/11

    DOI： 10.1016/j.jbiosc.2009.08.023 　

    ISSN： 1389-1723

  14. 89. Mode of action and cell response of jasmonate glucosides

    Nakamura Yoko, Hamamoto Shin, Inomata Sho, Mithofer Axel, Boland Wilhelm, Uozumi Nobuyuki, Ueda Minoru

    (44)　103-103　2009/10/06

    Publisher: The Japanese Society for Chemical Regulation of Plants

    ISSN： 0919-1887

    More details Close

    Samanea plants close their leaves in the evening, as if to sleep, and open them in the morning according to the circadian rhythm. Potassium β-D-glucopyranosyl-12-hydroxyjasmonate (1) was isolated as leaf-closing factor (LCF) of S. saman. We developed molecular probes consisting of modified LCF1 in order to identify its mode of action. We have already demonstrated that a specific binding protein is involved in the motor cell of S. saman. We synthesized natural-type photoaffinity probe and biologically inactive enantiomer-type probe. We utilized them for photoaffinity labeling of the target protein for LCF1. By using protoplasts of motor cell, we found membrane protein of 38kDa which strictly recognizes the stereochemistry of 1, and it is highly likely that the protein is the target protein for LCF 1. Recently, we observed that LCF shrank motor cell protoplasts prepared from S. saman. And comparing the results of several bioassay using glucosyl jasmonate-type LCF and jasmonic acid, it is also interesting that the mode of action of LCF is completely different with that of jasmonate.

  15. 2Ep20 Role of Na transporters, AtHKT1 and AtNHX1on Na tolerance in plants

    SUGIMOTO Yu, ISHIKAWA Atsushi, UOZUMI Nobuyuki

    21　65-65　2009

    Publisher: 日本生物工学会

  16. 2Ep21 Study on the regulation mechanism of Na-dependent K transporter Ktr system from Synechocystis :

    Zulkifli Lalu, UOZUMI Nobuyuki

    21　66-66　2009

    Publisher: 日本生物工学会

  17. 微生物用パッチクランプ法を用いた植物イオンチャネルの機能解析

    浜本晋, 魚住信之

    バイオサイエンスとインダストリー　66　(12)　671-673　2008/12/01

    Publisher: バイオサイエンスとインダストリー協会

    ISSN： 0914-8981

  18. 2P1-C21 On-chip cell manipulation systems : Part 8: On-chip cell observation system using micropattern of hydrogel

    ARAI Fumihito, MATSUMOTO Hideyuki, SIZYUKU Toshiaki, UOZUMI Nobuyuki

    2008　"2P1-C21(1)"-"2P1-C21(4)"　2008/06/06

    Publisher: The Japan Society of Mechanical Engineers

    More details Close

    We succeeded in making a long time observation system of individual cells using PDMS microchip which has a cage made of permeable membrane. The cage is made by photolithography and it has fine resolution. The height of the cage is about 2 μm. Optical tweezers is used to manipulate each individual cell. We transported cells into the cage using optical tweezers and changed solution around cells. The size change of each cell was monitored for long time stably. This system provides us a stable environment to observe cells in various conditions and it is useful to investigate unknown biological mechanisms of cells.

  19. 2S3p01 E. coli and yeast as expression system for plant K transport proteins

    UOZUMI Nobuyuki

    20　34-34　2008

    Publisher: 日本生物工学会

  20. 2Ia09 On-chip real-time observation of osmotic stress adaptation in cyanobactaria

    SHIJUKU Toshiaki, MATSUMOTO Hideyuki, AKAI Masaro, ARAI Fumihito, UOZUMI Nobuyuki

    20　211-211　2008

    Publisher: 日本生物工学会

  21. 2Aa16 Evaluation of tobacco K^+ channel in yeast tonoplast

    Hamamoto Shin, Mori Yasuo, Nakanishi Yoichi, Maeshima Masayoshi, Yabe Isamu, Uozumi Nobuyuki

    20　62-62　2008

    Publisher: 日本生物工学会

  22. 2E16-2 Functional analysis of the Na^+/H^+ antiporter(NhaS3) in Synechocystis PCC6803

    TSUNEKAWA Kenta, SHIJYUKU Toshiaki, KOJIMA Youichi, KOBAYASHI Hiroshi, NAKAMURA Tatsunosuke, KURODA Teruo, OMATA Tatsuo, Uozumi Nobuyuki

    19　123-123　2007

    Publisher: 日本生物工学会

  23. Localization and function of K(+)channels from tobacco cells

    Shin Hamamoto, Junichiro Marui, Ken Matsuoka, Tetsuro Mimura, Tsuyoshi Nakagawa, Yoshiyuki Murata, Yoichi Nakanishi, Masayoshi Maeshima, Isamu Yabe, Nobuyuki Uozumi

    PLANT AND CELL PHYSIOLOGY　48　S122-S122　2007

    ISSN： 0032-0781

  24. Topogenesis of conserved ion selective filter of K^+ channel and K^+ transporter

    UOZUMI Nobuyuki

    76　(5)　449-452　2004/05/25

    Publisher: 日本生化学会

    ISSN： 0037-1017

  25. Mechanosensitivity of G Protein-Activated Inwardly Rectifying K^+ (GIRK) Channels Depends on Channel-PIP_2 Interactions throughthe Activation of PKC

    Zhang Liyan, Lee Jong-kook, John Scott, Uozumi Nobuyuki, Kodama Itsuo

    Circulation journal : official journal of the Japanese Circulation Society　67　211-211　2003/03/01

    Publisher: Japanese Circulation Society

    ISSN： 1346-9843

  26. Mechanosensitivity of G protein-activated inwardly rectifying K+ (GIRK) channels depends on channel-PIP2 interactions through the activation of PKC

    LY Zhang, JK Lee, SA John, N Uozumi, Kodama, I

    BIOPHYSICAL JOURNAL　84　(2)　225A-225A　2003/02

    ISSN： 0006-3495

  27. Identification of residues essential for Na+/K+ selectivity in HKT-type K+ transporters

    P Maser, E Brendan, J Schroeder, K Yamada, S Oiki, E Bakker, S Goshima, Y Hosoo, N Uozumi

    PLANT AND CELL PHYSIOLOGY　43　S101-S101　2002

    ISSN： 0032-0781

  28. Structure and function of the Na+/K+ translocating AtHKT1 transporters from plants

    Nobuyuki Uozumi, Yasuhiro Kato, Nobuyuki Matsuda, Yoko Sato, Akifumi Mizutani

    Recent Res. Devel. Membrane Biol.　1　119-126　2002

  29. EXPRESSION PATTERN OF Na^+/K^+ TRANSPORTER (ATHKTI) IN ARABIDOPSIS :

    KUBO Masahiro, YAMAGAMI Mutsumi, YAMAGUCHI Katsushi, NISHIMURA Mikio, Schroeder J. I., MATSUOKA Ken, UOZUMI Nobuyuki

    Plant and cell physiology　42　s209　2001

    Publisher: Japanese Society of Plant Physiologists

    ISSN： 0032-0781

  30. STUDY OF MEMBRANE TOPOGENIC FUNCTION OF PLANT VOLTAGE-DEPENDENT POTASSIUM CHANNEL :

    SATO Yoko, SAKAGUCHI Masao, NAKAMURA Tatsunosuke, GOSHIMA Shinobu, UOZUMI Nobuyuki

    Plant and cell physiology　42　s209　2001

    Publisher: Japanese Society of Plant Physiologists

    ISSN： 0032-0781

  31. STUDY OF MEMBRANE TOPOGENESIS OF ARABIDOPSIS VOLTAGE-DEPENDENT POTASSIUM CHANNEL, KATl :

    SATO Yoko, SAKAGUCHI Masao, NAKAMURA Tatsunosuke, UOZUMI Nobuyuki

    Plant and cell physiology　41　s25　2000

    Publisher: Japanese Society of Plant Physiologists

    ISSN： 0032-0781

  32. PHENYLETHYLAMINE-INDUCED PRODUCTION OF ACTIVE OXYGEN SPECIES AND CYTOSOLIC Ca^<2+> ELEVATION IN TOBACCO SUSPENSION CULTURE

    KAWANO Tomonori, PINONTOAN Reinhard, UOZUMI Nobuyuki, MUTO Shoshi

    40　s81-s81　1999/03

    ISSN： 0032-0781

  33. CLONING AND CHARACTERIZATION OF A cDNA ENCODING A DUNALIELLA TERTIOLECTA Ca^<2+>-DEPENDENT PROTEIN KINASE (CDPK)

    PINONTOAN Reinhard, YUASA Takashi, ANDERCA Marinela I., MATSUOKA Takashi, UOZUMI Nobuyuki, MORI Hitoshi, MUTO Shoshi

    40　s78-s78　1999/03

    ISSN： 0032-0781

  34. SUGAR-INDUCED CYTOSOLIC CALCIUM TRANSIENT IN ARABIDOPSIS THALIANA

    FURUICHI Takuya, UOZUMI Nobuyuki, MUTO Shoshi

    40　s81-s81　1999/03

    ISSN： 0032-0781

  35. Current topics of cation transporters in animal, plant and bacteria

    T. Nakamura, N. Uozumi

    Tanpakushitsu kakusan koso. Protein, nucleic acid, enzyme　44　(13)　1988-1995　1999

    ISSN： 0039-9450

  36. Molecular biology of generation and transduction of calcium signal in photosynthesis functional control. (Ministry of Education)

    飯田秀利, 魚住信之, 鳥山尚志, 中川祐子

    植物個体における光合成機能統御の分子基盤 平成10年度研究成果報告書　1999

  37. Molecular biological studies of calcium signal generation and transduction involving with photosynthetic functional control. The Ministry of Education S.

    飯田秀利, 魚住信之, 松岡隆司, 鳥山尚志, 村田紀夫, 及川良一, 中川祐子, 荒川志穂

    植物個体における光合成機能統御の分子基盤 平成9年度　1998

  38. サリチル酸および類似化合物によるタバコ培養細胞の[Ca^<2+>]_C上昇 : 構造と生理活性の相関

    河野 智謙, 高橋 宏二, 魚住 信之, 武藤 尚志

    日本植物学会大会研究発表記録 = Proceedings of the annual meeting of the Botanical Society of Japan　61　221　1997/09

  39. サリチル酸はタバコBY-2細胞の一過的[Ca^<2+>]_<cyt>上昇を誘導する

    河野 智謙, 佐橋 信哉, 高橋 宏二, 魚住 信之, 武藤 尚志

    日本植物学会大会研究発表記録 = Proceedings of the annual meeting of the Botanical Society of Japan　60　236　1996/10

Show all ︎Show first 5

Books and Other Publications 44

  1. 植物の脂質修飾型キナーゼによるCa2+シグナリング制御

    斎藤俊也, 魚住信之

    細胞 ニューサイエンス社　2022

  2. 環境ストレスに対抗する植物イオンチャネル制御メカニズム

    佐藤奏音, 石丸泰寛, 魚住信之

    エヌ・ティー・エス社,　2022

  3. カリウム輸送とその制御機構

    斎藤俊也, 石丸泰寛, 魚住信之

    日本土壌肥料科学雑誌, 92, 2, 99-107　2021

  4. 植物における硫黄代謝と光合成制御

    辻井雅, 石丸泰寛, 魚住信之

    生化学, 93巻5号 643-650　2021

  5. 葉緑体の光合成活性にかかわるイオン輸送体の最近の知見と動向

    辻井雅, 魚住信之

    生化学 みにれびゅう　2020

  6. 微生物TRPチャネルの機能と役割

    魚住信之, 山下敦子

    医学のあゆみ 別冊 106-112 医歯薬出版 (2019年270巻の別冊版）　2020

  7. イオン輸送体

    魚住信之

    続・生物工学基礎講座 バイオよもやま話 96, 589-592 生物工学　2018

  8. 植物への遺伝子導入と遺伝子発現 化学便覧 応用化学編 第7版

    日本化学会 編／丸善出版　2014

  9. 植物の乾燥ストレス耐性に関わるK＋チャネルの制御機構

    浜本晋, 七谷圭, 魚住信之

    バイオサイエンスとインダストリー　2013

  10. 植物と微生物の駆動力と膜輸送体 動物の相違点と共通性

    浜本晋, 七谷圭, 佐藤陽子, 魚住信之

    化学と生物 50(2) 86-92　2012

  11. Structure-Function Correlates in Plant Ion Channels

    Uozumi, N, Deryer, I

    In Egelman, E. (ed.) Comprehensive Biophysics Volume 6 - Channel Proteins Elsevier BV Amsterdam, 234-245　2012

  12. Ishimaru, Y., Hamamoto, S., Uozumi N., and Ueda, M. Regulatory mechanism of plant nyctinastic movement: An ion channel-related plant behavior

    Ishimaru, Y, Hamamoto, S, Uozumi N, Ueda, M

    Plant Electrophysiology 125-142 In Volkov, A. G. (ed.) Spring-Verlag, Berlin Heidelberg　2012

  13. Potassium and sodium transporters: Improving salinity tolerance in plants.

    2012

  14. 植物のNa循環系と耐塩性

    魚住信之

    日本土壌肥料学雑誌 81 (5) p65-69　2011

  15. Comprehensive Biophysics Volume 6 - Channel Proteins

    Uozumi, N, Deryer I

    Elsevier BV Amsterdam　2011

  16. Ion channels and plant stress: past, present and future

    Uozumi, N, Schroeder J.I

    Springer-Verlag Berlin Heidelberg　2010

  17. 微生物用パッチクランプ法を用いた植物イオンチャネルの機能解析

    浜本晋, 魚住信之

    バイオサイエンスとインダストリー 66, 671-673　2008

  18. チャネルタンパク質の膜への組込み様式

    魚住信之

    2006/10

  19. 膜電位依存性Kチャネル

    魚住信之

    2005/10

  20. Characterization of potassium channels from Arabidopsis thaliana.

    Ibuki T, Tokida Y, Matsuda N, Czempinski K, Muller-Roeber B, Ona T, Uozumi N

    2004/10

  21. Large-scale production of hairy root

    Uozumi N

    2004/10

  22. K＋チャネルの膜挿入様式とイオン透過孔の保存性

    魚住信之

    2004/10

  23. 植物カリウムイオン輸送系と生理的役割。（植物の膜輸送システム ポンプ・トランスポーター・チャネル研究の新展開

    水谷昭文, 佐藤陽子, 松田信行, 加藤靖浩, 魚住信之

    2003/10

  24. Ｋ＋イオン輸送系の植物成長と塩感受性への役割 （植物の代謝コミュニケーション 植物分子生理学の新展開

    佐藤陽子, 水谷昭文, 魚住信之

    2003/10

  25. Structure and function of the Na+/K+ translocating AtHKT1 transporters from plants. Recent Research Developments in Membrane Biology.

    Nobuyuki Uozumi, Yasuhiro Kato, Nobuyuki Matsuda, Yoko Sato, Akifumi, Mizutani

    2002/10

  26. 植物のK+チャネル・K+トランスポーターはどこまでわかったか

    魚住信之

    2001/10

  27. Molecular mechanisms of potassium and sodium transport in plants

    Schroeder, J, Buschmann, P, Eckelman, B, Kim, E, Sussman, M, Uozumi, N, Maser, P

    In W. J. Horst, M. K. Schenk, A. Bürkert, N. Claassen, H. Flessa, W. B. Frommer, H. Goldbach, H. -W. Olfs, V. Römheld, B. Sattelmacher, U. Schmidhalter, S. Schubert, N. v. Wirén and L. Wittenmayer (ed.) Plant Nutrition 92, Developments in Plant and Soil Sciences, Springer Netherlands　2001

  28. Genetic transformation of Ajuga reptans.

    Tanaka, N, Uozumi, N, Kobayashi, T

    1999/10

  29. 動植物と細菌のカチオン輸送系の接点 蛋白質核酸酵素

    中村辰之介, 魚住信之

    1999/10

  30. 植物細胞の培養と再分化に関する工学的研究

    魚住信之

    1999/10

  31. 真核生物のイオン輸送体を大腸菌で解析する

    魚住信之

    1999/10

  32. Artificial seed production through hairy root regeneration.

    Uozumi, N, Kobayashi, T

    1997/10

  33. セロリ再分化植物体から誘導したカルスを用いた植物体再生系と再分化効率

    末原憲一郎, 後藤伸介, 魚住信之, 小林 猛

    1996/10

  34. 植物組織培養における計測 インテリジェント農業ー自動化・知識化のすすめー

    魚住信之, 小林, 猛

    1996/10

  35. Artificial seed production through encapsulation of hairy root and shoot tips.

    Uozumi, N, Kobayashi, T

    1995/10

  36. Application of hairy root and bioreactors

    Uozumi, N, Kobayashi, T

    1994/10

  37. 食品関連酵素 酵素工学

    小林 猛, 魚住信之

    1993/10

  38. Secretion of thermophilic bacterial cellobiohydrolase in Saccharomyces cerevisiae.

    Uozumi, N, Iijima, S, Kobayashi, T

    1993/10

  39. Functional roles of active-site residues of Bacillus polymyxa b-amylase. Enzyme Engineering XI,

    Uozumi, N

    1992/10

  40. Production of artificial seeds. pp. 270-273 In S. Furusaki, I. Endo, R. Matsuno (ed.) Biochemical Engineering for 2001,

    Kobayashi, T, Uozumi, N

    1992/10

  41. バイオ利用エネルギー変換

    小林 猛, 魚住信之

    1992/10

  42. 人工種子大量調製法の開発

    小林 猛, 魚住信之

    1992/10

  43. 植物の分化を司る遺伝子

    魚住信之

    1992/10

  44. 細胞膜上の分業別地図

    魚住信之

    1990/10

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Research Projects 29

  1. 「植物の養分吸収と循環系」総括班

    西澤 直子, 松岡 健, 藤原 徹, 魚住 信之

    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: 東京大学

    2005 - 2011

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    総括班は研究期間を通じて研究者間の連絡調整と研究支援、特定研究推進の方向性の検討と助言、他のプロジェクト研究との情報交換、本特定領域研究の広報活動等を行なってきたが、研究期間全体としての総括と成果のとりまとめを行う。また、領域メンバー間で共有可能なデータや材料も集まりつつある。これまでの全研究期間におけるデータの集約化を図ることにより、今後のこの研究分野の発展に資するものとして努めてきた。 平成21年度までで当初め研究期間を終了し、本年度においては、これまでの研究成果を取りまとめ、書籍等によって公表するとともに、関連研究者などに広く研究成果を公表することを目的としている。 平成23年3月11日、東日本大震災に付随して引き起こされた東電福島第一原子力発電所の重大な事故によって、大量の放射性降下物(fallout)が広域にわたって放出され、森林、農地、都市、海域に拡散した。農作物が放射性物質によって汚染され、食物連鎖を通して生物濃縮されつつある。農地土壌の汚染は、その土壌に栽培された農作物の汚染、食品の汚染につながった。土壌からの作物による放射性物質の吸収と可食部への移行、集積は、特定領域研究「植物の養分吸収と循環系」で扱ってきた対象である。そこで、研究班員の最新の成果をレビューした講座(日本土壌肥料学会誌に掲載)の再録と共に、放射性降下物に関する論文の復刻版を企画し、ニースレター追補版として発行した。発行後は特定領域班員だけでなく、日本学術会議シンポジウム等で無償で配布した。

  2. Study on plant Na and K transport system, which is involved in the formation of membrane potential and adaptation of osmolality

    UOZUMI Nobuyuki

    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

    2005 - 2009

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    The membrane transport system is closely involved in the formation of membrane potential and adaptation to the abiotic stress. We aimed the physiological role of the channels and the transporters which mediated Na and/or K permeation across the plasma membrane and thylakoid membrane. There are some difference in Na and K transport system between animal cells and plant cells. We have studied the structure and function of the Na/K transporters and K channels in plant cells and cyanobacteria which are an essential system for the controlling the solute flux across the membrane.

  3. Analysis of function of ion channel using giant bacteria and identification of membrane transporters

    UOZUMI Nobuyuki

    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

    2007 - 2008

  4. Molecular analysis of ion transporters in plant and cyanobacteria involved in response to high osmolality

    UOZUMI Nobuyuki

    Offer Organization: Japan Society for the Promotion of Science

    System: Grants-in-Aid for Scientific Research

    Category: Grant-in-Aid for Scientific Research (B)

    2005 - 2006

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    1. We have identified the localization of Arabidopsis Na transporter AtHKT1 in plant cells. The specificity of the anti-AtHKT1 antibodies was confirmed. Immunoelectron microscopy using the antibodies demonstrate that AtHKT1 is targeted to the plasma membrane in xylem parenchyma cells in leaves. 2. AtHKT1 disruption alleles caused large increases in the Na content of the xylem sap and conversely reduced the Na content of the phloem sap. The athkt1 mutant alleles had a smaller and inverse influence on the K content compared with the Na content to the xylem, suggesting that K transport may be indirectly affected. The expression of AtHKT1 was modulated not only by the concentrations of Na and K but also by the osmolality of non-ionic compounds. 3. The conserved Arg near the middle of the M2D segment was essential for the K^+ transport of KtrB. We propose the existence of the salt bridge(s) between the positive residues in the M2D and the conserved negative residues in the pore region, which reduces an electrostatic barrier against cation permeation caused by the positive residue(s) and that stabilizes the transporter configuration. 4. Mutation of a conserved His-157 in the second pore loop of KtrB, drastically reduced the activity of the K^+ transporter KtrABE system from Synechocystis PCC 6803. The result suggests that His-157 plays an essential role in the K^+ transport activity of the transporter system. 5. H transport activity of one of Synechocystis Na/H transporters was measured using the inverted membrane vesicle of E. coli expressing the protein. We have identified the charged residues essential for the K transport.

  5. Localization mechanisms of membrane proteins in the plant secretory pathway

    MATSUOKA Ken, TOYOOKA Kiminori, UOZUMI Noboyuki

    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: RIKEN

    2003 - 2005

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    In order to utilize higher ability of the glycan synthesis in plants and to produce valuable and bioactive glycans using plant cell culture or plant cultured roots, modified glycan syntheses should be localized to the Golgi apparatus, where most of the cell wall non-cellulose glycans synthesized. Plants also have metabolic pathways for the catabolism of complex compounds using membrane-anchored oxidoreductases. The industrial use of these enzymes requires high-level production of the proteins. Thus developing procedures to synthesize such membrane proteins with high quantity is necessary. Moreover, most of the stresses that prevent the growth of plants in non-optimum environment can overcome by the modulation of various transporters. Thus developing systems to engineer localizations and stabilities of integral membrane proteins including transporters are requested. At present, however, knowledge on the localization mechanism of membrane proteins are scarcer than the one of soluble proteins. Therefore, in this work we aimed to clarify the signals and mechanisms of the localization and degradation of membrane proteins in plant cells using cytochrome b5 and prolyl hydroxylase as major tools. In addition to this work we cloned several membrane transporter cDNAs from tobacco and analyzed their localization. As a result, the following results were obtained. 1)Prolylhydroxylase is a typeII membrane protein localizing to the Golgi apparatus. The Golgi localization or efficient export from the ER requires basic residues in the cytoplasmic tail. 2)Fusion protein of cytochrome b5 and a red fluorescent protein form stable protein aggregate in the cell. Formation of the aggregate depends on the tetrameric nature of RFP. 3)From tobacco BY-2 cells several transporter and other membrane protein cDNAs were cloned and localization of some of them are characterized.

  6. Molecular mechanism of flexible development and differentiation in plants

    MATSUOKA Makoto, HATTORI Tsukaho, SAKAGAMI Youji, MAESHIMA Masayoshi, UOZUMI Nobuyuki, MIZUNO Takeshi

    Offer Organization: Japan Society for the Promotion of Science

    System: Grants-in-Aid for Scientific Research

    Category: Grant-in-Aid for COE Research

    Institution: NAGOYA UNIVERSITY

    2001 - 2005

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    ○Study on molecular mechanism of plant growth We identified a receptor of gibberellin (GA). which is an important growth phytohormone. The GAreceptor interacts with bioactive GAs at a reasonable concentration. The binding GA to the receptor induces the interaction between GID1 and a DELLA protein, which functions as a negative factor of GA signaling. Cytokinin is another growth phytohomone. We identified the three genes encoding cytokinin receptors and investigated their relevant in planta functions. We also revealed that a grain number in rice is controlled by the level of cytokinin in the floral meristems. Using the graining number gene, we increased the grain number of Koshihikari, the top rice leading variety of Japan. We identified a set of components (or genes), named PRRs, which play coordinately essential roles within (or dose to) the Arabidopsis circadian clock that is important for the control of the flowering time in plants. We also identified a receptor of plant peptide hormone, pktosulfokine, discovered by us. ○Study on molecular mechanism of plant response to physical environments. We found characteristic properties of five cation/H+ exchangers of rice. Ktr/HKTtype transporters were also found to be essential for the adaptation of plants and bacteria to salinity stress and high osmolality We also revealed that nitrate transporter genes are activated by multiple mechanisms involving nitrate and/or nitrite, and also that glutamine was shown to regulate nitrate transport activity at both the transcriptional and post-translational levels. For understanding of signal transduction of responses to hyperosmotic stress, we have identified an SnRK2 interacting protein that would function as a scaffold for signaling components.

  7. 原核生物・真核生物のイオン輸送体を統制する構造の探索

    魚住 信之

    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: 名古屋大学

    2004 - 2004

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    1 ラン藻(Synechocystis sp. strain PCC 6803)のKtr系はCCCPの添加で阻害され、H駆動力のない変異株に発現させると輸送活性が検出できなかったことから、H駆動力がKtr系輸送体に関与することが示唆された。また、ラン藻に高浸透圧ストレスを与えるとKの流出が起こり、そのK流出を補うKの取込みに関与して、浸透圧変化に適応するのに必須なK取込み系として機能していることが明らかとなった。らん藻のHKT/Trk/Ktr系Na/Kトランスポーターにおいて、他の生物のホモログ輸送体からは見いだされていない細胞質性因子KtrAとKtrEをHisタグ融合蛋白質として大腸菌で発現させて精製した。 2 ラン藻・シロイヌナズナ・小麦のNa/Kトランスポーター(KtrABE,AtHKT1,wHKT1)は8回の膜貫通構造をとる。第8番目の膜貫通領域に存在する正電荷アミノ酸を他のアミノ酸に置換してイオン選択性への関与を検討した。野性株のArgからLysへの置換はイオン透過性が保持されたが、負電荷アミノ酸や極性のないアミノ酸への置換はイオン透過性が失われた。この正電荷アミノ酸は本輸送体の機能に重要であることが分かった。 3 小胞体膜挿入系を用いてShaker型Kチャネルの膜挿入機構を検討したところ、植物のKチャネルと比較して膜電位センサー以外の膜貫通領域の挿入機構は同様であり、膜電位センサーを形成する第3番目の膜貫通領域は自律的に膜へ挿入することが明らかとなった。 4 ラン藻のNa/Hトランスポーターのうち他の生物では見いだすことができない特徴的な配列を保持した機能不明のトランスポーターを大腸菌発現系で機能発現することが分かり、Na排出活性があることを確認した。植物の報告されているNa/Hトランスポーターと機能の観点からも高いことが分かった。

  8. Molecular analysis of plant Na and K transporter involved in response to environmental stress

    UOZUMI Nobuyuki

    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: Nagoya University

    2003 - 2004

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    1 The Ktr-system in Synechocystis sp.PCC 6803 was found to be Na-activated K transporter. The K transport activity was inhibited by CCCP and was not found in the mutant strain which has a mutation of the respiratory chain system. These indicate that proton motive force is involved in the Ktr-mediated K transport. Hyperosmotic stress led initially to loss of net K from the cells. The ktrB mutant shocked with sorbitol failed to reaccumulate K up to its original level. The K uptake via KtrABE palys a crucial role in the early phase of cell turgor regulation after hyperosmotic shock. 2 Bacterial Ktr system and plant HKT-system has eight transmembrane topology. The positive residue in the eighth transmembrane segment was replaced with other amino acids. The conversion from Arg to Lys sustained the K uptake activity but that to negative or non-polar amino acids gave loss of the function. The positive residue is important for the activity. 3 KUP-type transporter was not inserted into the rough microsomal membrane. Then the transporter was expressed in E.coli and their membrane structure was determined. 4 Some of plant K channels has the putative Ca binding sites. The mutation in the region inhibited the K uptake activity. The region is important for the transport activity. 5 Arabidopsis AtHKT1 transporter was expressed in the vascular tissuels in the aerial part of the plant. The null mutant showed that Na accumulated in the xylem tissue and Na was not supplied into the phloem tissue. AtHKT1 is one of the components for Na recirculation of plant.

  9. 原核細胞イオン輸送系の解析に基づく種を超えた基本構造の探策

    魚住 信之

    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: 名古屋大学

    2003 - 2003

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    らん藻Ktr系の破壊株のイオン輸送機能を野生株と比較した。らん藻の破壊株のK^+輸送は減少していることが分かった。また、本輸送系はNa^+に強い依存性があるが、Na^+の輸送活性はほとんど見いだせなかった。また、pHは中性付近で最も輸送活性が高いが、プロトンイオノフォアの添加で輸送活性が阻害されたことから、プロトン駆動力が必須であることが明らかとなった。一方、Ktr破壊株で検討した培養条件で同じ表現系を示すらん藻の二成分系変異株をスクリーニングしたが、Ktr系の破壊株で阻害を受ける二成分系変異株は取得できなった。 Ktr/HKT系のイオン選択孔の中には塩基性アミノ酸(Arg)が存在する。ArgをLysへの置換はK^+輸送活性に影響を与えなかった。一方、AlaやGluへの置換はK^+輸送活性を減少させた。本残基がNa^+選択性にも関与することが示唆された。 シロイナズナおよび大腸菌KUPの膜貫通構造をCystein scanning法で決定する。既に蛋白質内に存在する4っのCysをSerに置換してもK^+輸送活性を維持することを確認している。膜貫通領域の前後にCysを導入し、スフェロプラストを用いて膜透過性NEM-biotinmaleimideと膜非透過性MTSETの2つのSH試薬を用いて膜トポロジーを決定したした。先にアルカリフォスファターゼ融合法で決定したシロイナズナAtKUP1のトポロジーと比較したところ異なる箇所が見出された。

  10. イオン輸送系膜蛋白質の構造と機能の解析法の構築

    魚住 信之

    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: 名古屋大学

    2002 - 2002

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    KUP系K^+輸送体の膜貫通構造の決定と決定方法の検討:Cysに特異的な修飾剤を用いて、大腸菌で発現しているKUP系輸送系のtopologyの決定を行った。その結果、明確な膜貫通構造を形成する部位と曖昧な領域が明確になった。アルカリホスファーターゼ(PhoA)を膜蛋白質に融合した結果はCys scanning法を用いて決定した結果と異なった。PhoA融合決定法はCys scanning法と比較して精度が低いことが示めされた。 HKT系トランスポーターのイオン選択残基とチャネルとの共通性:Na^+とK^+のイオン選択性に関与するアミノ酸に部位特異的変異を導入して、アフリカツメガエル卵母細胞発現系を用いた電気生理学的測定と酵母変異株を行いてイオン透過性を検討した。この結果、4つのイオン選択孔に存在するイオン選択性に重要な影響を与える残基が4カ所ともにGlyの時(小麦HKT1等の場合)は、K^+を輸送しSerの場合(AtHKT1等の場合)はK^+輸送活性がないことが分かった。このことは、Ktr/Trk/HKT系トランスポーターとK^+チャネルの構造の類似性を示しており、Ktr/Trk/HKT系トランスポーターが単独MPM-K^+チャネルから進化して形成されたことを強く示している。 藍藻のHKTホモログ:藍藻(Synechocystis PCC6803)のHKT系輸送体のK^+取り込み活性が測定され、さらにNa^+によってK^+輸送活性が活性化されることがK^+取り込み系欠損およびNa^+感受性の大腸菌変異株を用いた発現系で解析したところ分かった。 膜依存性イオンチャネルの膜への組込み機構:植物から動物に存在する膜電位依存性イオンチャネルの膜電位センサーを担う構造の形成を解析した。2つの膜貫通領域が同時に挿入される様式であることが分かった。これはまだ知られていない新規の挿入機構である。

  11. Physiological function of Na^+/K^+ transporters from bacteria and plants

    UOZUMI Nobuyuki

    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: NAGOYA UNIVERSITY

    2001 - 2002

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    1. AtHKTl from Arabidopsis (AtHKTl) was found to form 4-fold membrane-pore-membrane structure. 2. It is found that the P-loop of HKT transporters show identity only in one of glycine residues that mediates K^+ selectivity. HKT transporters share a common ancestor transporter with 4-loops. 3. KATl shares typical similarity with other animal and plant K^+ channels. The voltage-sensing segments of S3 and S4 in KATl are posttranslationally integrated into the membrane only when specific interation occurs between them. The insertion process is a newly recognized second type of the membrane protein integration process. 4. The aspartate in the transmembrane of Na^+/H^+ antiporter from a bacteria involves in H^+ permeation but not Na^+ permeation. 5. The AtHKTl was expressed in vascular tissue in plant. 6. AtHKTl plays a role in the recirculation of Na^+ in plant body. 7. The K^+ permeation of HKT-type transporter in Synechocystis PCC6803 was measured. K^+ transport activity was activated by Na^+ using E. coli expression system.

  12. 大腸菌を用いた真核生物のイオン輸送系膜蛋白質の解析

    魚住 信之

    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: 名古屋大学

    2001 - 2002

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    (1)AtKUP1(KUP系)のトポロジーを大腸菌発現系を用いて決定したところ、意外にも4回膜貫通構造であった。(疎水領域は13カ所存在する)。次にこの結果の真偽を検証し、大腸菌の発現系の有効性を評価するために、検証の必要な箇所をイヌの膜小胞や動物細胞HEK293を宿主とする発現系を用いた糖鎖修飾実験を行い大腸菌の結果と比較した。その結果、大腸菌発現系と異なる結果が得られた貫通領域が見つかった。 (2)HKT系(AtHKT1)のトポロジーから推定されるイオン透過孔に存在する特徴的なアミノ酸を置換し、Na^+とK^+の選択部位を決定したところ、AtHKT1のSer68とMet69がGly-Leuに変換されることにより、K^+透過選択性に変わることが明らかとなった。Na^+とK^+の選択部位の決定は本輸送系の本質的でアイデアルばかりではなく、あらゆるK^+の輸送系に必須のイオン選択孔の構造の一般化に重要な知見となる。 (3)Synechocystis sp. PCC6803のKtr系を大腸菌に発現させて解析したところ、KtrABCの3つのサブユニットでK^+の輸送を行うことを明らかにした。またNa^+によって輸送活性が大きくなることを示した。 (4)K^+チャネル(KAT1)の親水性膜貫通領域はpost-translationalに小胞体膜に挿入されることを示し、以前の大腸菌で得られた結果と一致した。親水性膜貫通領域は隣の親水性膜貫通領域と相互作用をして生体膜に移行することがあることを見出した。 (5)植物にも存在する細菌Na^+/H^+antiporterの膜貫通領域に存在するAspはH^+の透過には関与するがNa^+の透過には関与しないことを示した。

  13. 大腸菌を外来遺伝子発現宿主とする膜蛋白質解析系の構築

    魚住 信之

    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: 名古屋大学

    2000 - 2000

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    我々は既に大腸菌を用いた真核生物由来膜蛋白質の活性発現と構造決定が可能であることを示しており、本研究では構造と機能を解析することを目標に以下の課題を進めた。1 Arabidopsis由来のAtKUP1(KUP系)の輸送体トポロジーをアルカリフォスファターゼ遺伝子融合法で大腸菌発現系を用いて決定した。13ヶ所存在する疎水領域のうち7ヶ所は膜貫通構造を形成することが明らかとなった。高い疎水領域であるのにもかかわらず膜に貫通していない部分が存在した。この領域は一部が膜に埋まってイオン輸送孔を形成している可能性が推定された。2 我々は既に植物AtHKT1(HKT系)のトポロジーを大腸菌発現系を用いて決定している。この結果が真核細胞でも同様であるかを明らかにするために、FLAG抗体認識部位を5カ所、糖鎖修飾部位を7カ所創出した後、動物の発現系およびイヌの膵臓由来の膜小胞を用いて検討した。この結果、大腸菌で決定したトポロジーとすべての点で一致しAtHKT1は8回膜貫通構造を有すること証明した。この決定したAtHKT1の膜貫通構造は他のグループが報告している10回貫通構造とは異なる結果となった。我々の構造モデルは複数の方法で導き出したことから信憑性がより高いと考えている。3 AtHKT1はアフリカツメガエル卵母細胞では検出できないK^+輸送能が大腸菌発現系では観察される。この理由としてAtHKT1に存在する一カ所のN型糖鎖修飾が関係している可能性がある。糖鎖修飾機能をもつ真核細胞発現系とそれを持たない大腸菌発現の差異がイオン選択性に影響を与えている可能性を検討した。糖鎖非修飾に残基置換したAtHKT1変異蛋白質を卵母細胞で発現させてイオン電流を膜電位固定法で調べたところ糖鎖修飾はイオン透過性には関与しないことがわかった。大腸菌は酵母や動物細胞よりもサイズが小さいため、K^+の要求性が小さくてAtHKT1は大腸菌のK^+取込み活性欠損変異を相補できたと考えている。酵母や動物の膜蛋白質発現系を補う別の蛋白質解析系としても有効であることを示すことができた。

  14. 真核表層膜蛋白質の大賜菌活性発現系の有効性の検討

    魚住 信之

    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: 名古屋大学

    2000 - 2000

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    1)植物のAtKUP1は大腸菌で活性発現することはすでに報告している。植物KUP系の輸送体トポロジーをPhoA融合法で大腸菌発現系を用いて決定した。13ヶ所存在する疎水領域のうち7ヶ所は膜貫通構造を形成することが明らかとなった。高い疎水領域であるのにもかかわらず膜に貫通していない部分が存在した。この領域は一部が膜に埋まっている可能性が考えられ、さらにイオン輸送孔を形成している可能性がある。限定したこの部分をさらに詳細に検討する必要がある。 2)抗体認識部位をAtHKT1に導入し動物の発現系で検討した。同時に糖鎖修飾部位も創出して小胞体膜を用いて膜領域の配向性を調べた。FLAG抗体認識部位を5カ所、糖鎖修飾部位を7カ所検討したところ大腸菌で決定したトポロジーとすべての点で一致した。この結果、大腸菌発現系、抗体認識法による動物細胞の発現系および小胞体膜を用いた糖鎖修飾法によってAtHKT1は8回膜貫通構造を有すること証明した。この決定したAtHKT1の膜貫通構造は他のグループが報告されている構造とは異なる結果となった。我々の構造モデルは複数の方法で導き出したことから信憑性がより高いと考えている。この膜貫通構造の決定に基づいてイオン選択孔の位置の特定を行うことが可能となり、現在そのイオン選択孔を解析している。 3)AtHKT1には一カ所N型糖鎖修飾が予想される残基が存在する。イオン透過孔に面する箇所の糖鎖修飾の有無がK+Na+のイオン選択性に影響を与えている可能性がある。糖鎖修飾機能をもつ真核細胞発現系とそれを持たない大腸菌発現系の差異を検討した。糖鎖非修飾に残基置換したAtHKT1変異蛋白質を作成して卵母細胞でイオン電流を膜電位固定法で調べたところ糖鎖修飾はイオン透過性には関与しないことがわかった。大腸菌と真核細胞の蛋白質への修飾の差異の一つとして糖鎖修飾の有無があるが、本膜蛋白質ではその影響ではなかった。大腸菌は酵母や動物細胞よりもサイズが小さいため、K+の要求性が小さいことからAtHKT1は大腸菌においてK+の取り込み活性変異を相補したと考えている。酵母や動物の膜蛋白質発現系を補うもう一つの蛋白質発現系として有効であると考えられる。

  15. Study on function of the diverse K^+ transporters from plants

    UOZUMI Nobuyuki

    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: NAGOYA UNIVERSITY

    1999 - 2000

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    1. A cDNA homologous to HKT1 from Arabidopsis (AtHKT1) and the characterization of its mode of ion transport in heterologous systems. The AtHKT1 mediates inward Na^+ currents in Xenopous laevis oocytes and Na^+ uptake in Saccharomyces cerevisiae, and K^+ uptake in Escherichia coli. 2. Independent experiments using as E.coli-expression system and PhoA reporter enzyme fusions, glycosylation reactions in a eukaryotic cell free system and HEK293 transfectants and immunofluorescence detection showed that AtHKT1 contains eight transmembrane-spanning segments. Our model is a more accurate topology than the model proposed by other groups. 3. The wild type AtHKT1 possesses the N-glycosylation site. An engineered unglycosylated protein variant mediated Na^+ currents in Xenopous laevis oocytes without loss of monovalent cation selectivity just as the wild type protein, indicating that glycosylation is not essential for either the expression of AtHKT1 in the plasma membrane of the Na^+ translocation activity of AtHKT1. 4. Although the wild-type plant hyperpolarization-activating K^+ channel, KAT1, was insensitive to external Na^+, the mutanl channels, T256Q and T256E, were significantly depressed by Na^+ with their apparent dissociation constants. At the extreme hyperpolarization the blocking was relieved significantly in the T256E.The mutation at position 256 within the pore helix rearranged the selectivity filter and allow Na^+ to penetrate into the pore. 5. When extracts from abscisic acid-treating Vicia fava guard cell were added into the carboxyl-terminus of an inward-rectifying K^+ channel from Arabidopsis KAT1, phosphorylation of the peptides were observed. 6. The topology of K^+ transporter, AtKUP1, were determined by PhoA fusion approach in E.coli gene expression system. We identified seven transmembrane segments in the AtKUP1.

  16. カルシウムシグナルによる光合成統御の器官間コミュニケーション

    鳥山 尚志, 魚住 信之

    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: 名古屋大学

    1999 - 1999

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    エクオリン導入アラビドプシス植物体をモデル植物体として用いて,糖シグナルに応答した[Ca^<2+>]_c上昇をCCDカメラと画像解析装置によってモニターした。また,^<14>C-蔗糖の移動と,蔗糖処理による植物体表面電位の変化を測定した。蔗糖を与えない培地で生育したエクオリン導入アラビドプシス植物体の根部に糖を与え,下位葉から上位葉へと経時的に移動する発光を,2分の時間分解能で解析した。発光の移動と14C-蔗糖の移動はほぼ一致した。また,植物体表面電位変化の移動も発光とほぼ一致した。表面電位の変化は,シンク状態にある細胞がH^+/蔗糖共輸送体によって通導組織から蔗糖を取り込み,その際にH^+を同時に取り込むために,シンク細胞で脱分極が起こったことを反映していると推測される。脱分極がおそらく電位依存性Ca^<2+>透過性チャネルを活性化して,細胞外からCa^<2+>を流入させ[Ca^<2+>]_c上昇をもたらしたものと推測される。[Ca^<2+>]_c上昇は,Ca^<2+>-依存性プロテインキナーゼなどのCa^<2+>依存性酵素の活性化を介して,同化糖の貯蔵に必要な酵素系を構成する酵素遺伝子の発現を誘導すると考えられる。 出芽酵母の仮想的カルシウムチャネルCCHl欠損を相補する遺伝子としてモノアミンオキシダーゼ(MAO)ホモログを得た。MAOの基質フェニルエチルアミン(PEA)をエクオリン導入midl酵母に与えたところ,細胞外Ca^<2+>の流入による[Ca^<2+>]_c上昇が起こった。PEAによるmidl酵母における[Ca^<2+>]_c上昇は,Ca^<2+>がMidl以外のチャネルか輸送体によって取り込まれたことを示している。MAOの反応生成物の一つはH_2O_2であり,これが引き金として働いて細胞内へのCa^<2+>流入を引き起こした可能性がある。

  17. 大腸菌を利用した真核細胞由来イオン輸送膜蛋白質の活性発現系の開発

    魚住 信之, 鳥山 尚志

    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: 名古屋大学

    1999 - 1999

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    1小麦Na^+K^+トランスポーター(wHKT1)のホモログ遺伝子をArabidopsisからクローン化して(AtHKT1)大腸菌で発現させた。wHKT1は大腸菌のK^+取り込み変異を相補しなかったが、AtHKT1は相補した。wHKT1が活性発現しなかった理由は明らかではない。次に、PhoAを用いたトポロジー解析を行った。更に、N末端についてはエピトープを導入して膜の内外について検討した。両者の方法からおおよそのトポロジー構造は決定できた。 2マウスCa^<2+>チャネルの膜貫通構造を大腸菌PhoA法によって一部を決定した。 3AtHKT1が発現した大腸菌のK^+取り込み変異株のK^+取り込み活性測定を原子吸光分析法によって測定した。この結果、AtHKT1が発現する大腸菌と相補性を示さない大腸菌のK^+取り込み速度を比較した場合、Km値はほぼ同一であったがVmaxが1.7倍程度大きくなっていた。一方、昨年度、植物K^+チャネル(KAT1およびAKT2)はK^+取り込み能を欠損した大腸菌を相補することを見いだしたが、K^+の取り込みは測定できなかった。パッチクランプ測定可能な大きさに大腸菌を巨大化し、大腸菌の細胞膜を対象に電気生理学的手法でK^+電流を測定することを試みた。K^+電流が観察されたが、今後、本電流はKAT1由来であることを数多くの実験で検証する必要があると考えている。

  18. カルシウムシグナルによる光合成統御の器官間コミュニケーション

    鳥山 尚志, 魚住 信之

    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: 名古屋大学

    1998 - 1998

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    アポエクオリン導入アラビドプシス植物体にセレンテラジンを与え,植物個体全体にエクオリンが構成させ,糖に応答したサイトソルCa^<2+>上昇を反映したエクオリン発光をVIMカメラを用いて測定した。自養栄養的に生育した植物体の根部に糖を与えると,下位葉から上位葉へと経時的に移動する発光が観察された。蔗糖を与えて生育した植物体では,このような発光は観察されなかった。先にタバコ培養細胞で観察された糖飢餓細胞が糖に応答した観測と対応した結果であり,器官間の糖シグナル伝達に細胞内カルシウムシグナリングが関与していることを示している。 前年度に取得したトウモロコシCa^<2+>/H+アンチポータcDNAは,出芽酵母Ca^<2+>/H^+アンチポータ遺伝子破壊株の高カルシウム培地での生育を回復させた。さらに遺伝子破壊株の相補試験の結果は,この輸送体はCa^<2+>以外にMg^<2+>やNa^+,K^+なども輸送することを示唆した。 出芽酵母のSOC1は動物の"Capasitative"あるいは"Store-operated"カルシウムチャネル類似のチャネルをコードする遺伝子であると想定されている。そこでアラビドプシスcDNAライブラリーからsoc1を相補する遺伝子の単離を試みた。相補する遺伝子は複数得られたものの,それらの塩基配列はいずれも膜蛋白をコードする遺伝子ではないことを示した。次に,既にデータベースに登録されているイオンチャネルのホモログが,Ca^<2+>透過能をもつ可能性を検討した。出芽酵母のカルシウムチャネルと想定されるMID1を欠損する変異体は,低Ca^<2+>培地中の生育が著しく低い。この変異体にアラビドプシスのデータベースに登録されているイオンチャネルホモログ遺伝子の一つを導入することによって,低Ca^<2+>培地中での生育が回復した。このことはこれがCa^<2+>透過能をもつことを支持している。 ^<45>Ca^<2+>の取り込み能,イオン選択性を検討中である。

  19. 大腸菌を用いた真核生物標的膜蛋白質の活性発現系の構築とイオン輸送系の改変

    魚住 信之, 鳥山 尚志

    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: 名古屋大学

    1998 - 1998

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    真核生物標的膜蛋白質の表面提示と活性発現系として大腸菌を利用することは、他の真核培養細胞で発現させる場合と比較して、操作性、経済性の点で有効であり、実験労力の低減を可能にする。現在まで難しいと考えられてきた単細胞生物の細菌に真核生物の膜蛋白質を容易に活性発現することが可能となれば、新規の発現系の開発として認知され真核膜蛋白質の分子デザインの研究に大きく寄与する。本研究から以下の成果を得ることができた。 1、 高等植物のイオン輸送体(KAT1,AKT2,AtKUP1)をK^+取り込み系が変異している大腸菌において活性発現を試みたところ、KAT1,AKT2,AtKUP1はK^+取り込み能を欠損した大腸菌を相補することを見いだした。K^+の初期濃度7.6-10.4mMの範囲では、対照のプラスミドを導入した大腸菌は増殖しなかった。また、イオン孔のアミノ酸が変異しているKAT1は機能しないことが分かっていたが、予想通り大腸菌においてもK^+輸送の欠損変異を相補しなかった。 2、 Shaker型K^+チャネルKAT1のトポロジーの決定を行った。疎水性プロットからKAT1には8箇所の疎水性領域(S1,S2、S3,S4、S5,H5,S6,S7)が存在する。推定膜貫通領域の前後に細胞外で活性を示すPhoAを連結した8種類のプラスミドを作成した。解析したところ従来から推定されていS1,S2,S3,S4,S5,S6は膜貫通領域であること、孔を形成するH5と疎水性の度合いが強いS7は膜を貫通していないことが明らかとなった。 真核生物由来の膜蛋白質が大腸菌で機能を持って発現することを証明した。さらに、構造と機能の解析が可能であることを示した。

  20. 高等植物K^+輸送の分子機構の解明

    魚住 信之

    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: 名古屋大学

    1997 - 1998

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    1 Arabidopsisから単離した2種類のKAT1およびAKT2は4量体で形成するK^+チャネルと考えられているがKAT1とAKT2が単独つまりホモ体で機能するのかヘテロ体でも機能するのかが明らかとなっていない。そこで、卵母細胞に両K^+チャネルを発現させて機能を調べた。KAT1のイオン透過孔を変異させたKAT1-T256RはKAT1またはAKT2を共発現させるとK^+チャネル活性を抑制した。このことは、KAT1とAKT2は互いに親和的相互作用を行うことを示している。N末またはC末の細胞質に突き出た箇所が重要であることが示唆される。 2 内向き整流性Shaker型K^+チャネルKAT1のトポロジーの決定を行った。疎水性プロットからKAT1には8箇所の疎水性領域(S1,S2,S3,S4,S5,H5,S6,S7)が存在する。推定膜貫通領域の前後に細胞外で活性を示すPhoAを連結した8種類のプラスミドを作成した。解析したところ従来から推定されていS1,S2、S3,S4,S5,S6は膜貫通領域であること、孔を形成するH5と疎水性の度合いが強いS7は膜を貫通していないことが明らかとなった。 3 K^+トランスポータ(AtHKT1)を酵母にて発現させたところNa^+は輸送しないでK^+を輸送しないことが分かった。また、新規K^+トランスポータ(AtKUP1)をArabidopsisからクローン化しK^+輸送活性を確認した。

  21. カルシウムシグナルによる光合成統御の器官間コミュニケーション

    鳥山 尚志, 魚住 信之

    Offer Organization: 日本学術振興会

    System: 科学研究費助成事業

    Category: 重点領域研究

    Institution: 名古屋大学

    1997 - 1997

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    カルシウム依存性発光タンパク質エクオリンを形質導入したタバコ培養細胞BY-2を利用して,糖シグナルに対応した細胞礎質カルシウムイオン濃度([Ca^<2+>]c)上昇を測定した。対数増殖期の細胞では,高濃度の糖を与えても[Ca^<2+>]c上昇は観察されなかった。定常増殖期の細胞では高濃度の糖によって[Ca^<2+>]c上昇が起こった。対数増殖期の細胞でも蔗糖飢餓状態にすると糖による[Ca^<2+>]c上昇が観察された。定常増殖期の細胞でも蔗糖飢餓処理により[Ca^<2+>]c上昇が顕著になった。これらの結果はシンク的状態の細胞が糖に対応することを示唆している。[Ca^<2+>]c上昇は蔗糖以外にマルトース,グルコース,フラクトースなどによっても誘導された。蔗糖と同じ浸透圧変化を引き起こす濃度のNaClやKClはわずかしか[Ca^<2+>]c上昇を引き起こさなかった。糖誘導[Ca^<2+>]c上昇は培地中のCa^<2+>を除くと起こらなくなった。また,Ca^<2+>拮抗剤La^<3+>によって阻害された。これらのことは[Ca^<2+>]c上昇は細胞外のCa^<2+>が流入したことを示唆している。プロテインキナーゼ阻害剤K-252aやスタウロスポリンは糖誘導[Ca^<2+>]c上昇を阻害した。このことは[Ca^<2+>]c上昇にタンパク質リン酸化が関与していることを示している。事実,インゲルプロテインキナーゼアッッセイによりプロテインキナーゼ活性上昇が明らかになった。 既知のCa^<2+>/H^+アンチポ-タの塩基配列を利用してPCR法により,トウモロコシcDNAライブラリーをスクリーニングし,Ca^<2+>/H^+アンチポ-タの全長をコードしていると考えられる遺伝子を取得した。410残基のアミノ酸をコードする遺伝子で,遺伝子産物は既知のCa^<2+>/H^+アンチポ-タ遺伝子と同様に,11回膜を貫通する構造を持つと予測された。このCa^<2+>/H^+アンチポ-タの細胞内分布を知る目的で,親水性領域のペプチド断片を大腸菌で生産してウサギを免疫して抗体を産生することを試みている。

  22. Mass production of regenerated plants by artificial seed system

    KOBAYASHI Takeshi, UOZUMI Nobuyuki, HONDA Hiroyuki

    Offer Organization: Japan Society for the Promotion of Science

    System: Grants-in-Aid for Scientific Research

    Category: Grant-in-Aid for Scientific Research (A)

    Institution: Nagoya University

    1994 - 1996

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    Efficient prodution system of adventive embryo, automatic selection system of adventive embryo and artificial seed system from hairy roots were studied extensively. Celery embryos and plantlets were found to be selectively released in a culture of immobilized Ca-alginate gel beads in which celery callus was entrapped under regeneration conditions. Repeated batch culture with 5 mM CaCl_2 provided long-term(more than 154d) embryo and plantlet production without gel beads disruption. Productivity of plantlets in the immobilized cell culture was 2,2-fold as high as that in the suspension culture. An efficient plantlet production from horseradish hairy roots was studied to apply the hairy roots for artificial seed. To make the hairy root fragments, a blender was applied after the root growth. A fragmentation time of 30 s gave the best result on the plantlet formation. Treatment of the hairy roots with naphthaleneacetic acid(1.0 mg/l) made the culture time shorter and increased plantlet productivity. Treatment with kinetin at 0.1 mg/l after fragmentation yielded the highest number of plantlets. Ajuga hairy root in which the beta-glucuronidase(GUS)gene was introduced under control of the tomato ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit (rbcS3B) promoter was constructed by the Agrobacterium rhizogenes-mediated cotansformation. The GUS-transformed hairy root could also be efficiently regenerated into plantlets through this procedure. GUS activity was detected in leaf tissue of the regenerated plants. For plant regeneration from fragmented tips of horseradish hairy root, a shaking-vessel type bioreactor was used. Number of regenerated plantlet depended on a rotational spped, and the maximum number was obtained at 120 rpm. It was concluded that the shaking-vessel type bioreactor was suitable for mass-production of regenerated plantlet.

  23. 動物細胞の固定化及び培養環境の制御による有用物質の効率的生産

    小林 猛, 水谷 悟, 魚住 信之, 本多 裕之

    Offer Organization: 日本学術振興会

    System: 科学研究費助成事業

    Category: 試験研究(B)

    Institution: 名古屋大学

    1993 - 1994

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    申請者はほぼ当初の計画通りに研究を行い、以下の実績を得た。 1.一段階の簡便な操作で長期培養可能な強度を持つ担体として光架橋性樹脂BIXを開発した。三種類の固定化方法により浮遊性と付着性の動物細胞両方を固定化することができ、担体は強度の面で優れ、細胞毒性がなく、細胞増殖も良好で、特にゼラチン粒子を添加することにより作成されたポーラスBIXでは栄養分の供給と酸素通気が顕著に改善されることがわかった。 2.多孔性セルロース担体に正電荷を持たせることによって、壁付着性細胞だけでなく、浮遊性細胞の固定化が可能となった。L-929細胞を多孔性セルロース担体に固定化してエリスロポエチン(EPO)の連続生産を行った。インナーループ型バイオリアクターが良い結果を与え、50日以上にわたって高いEPOの連続生産が可能であった。 3.溶存酸素は動物細胞培養における重要な制御因子である。我々は溶存酸素を一定に制御できるシステムを用い、DOを0.5から28mg/Lの範囲で血清入培地と無血清培地両方にて調べたところ、最適な細胞増殖は今まで報告してきた通りの5mg/Lであったが、EPOやティシュープラスミノーゲンアクチベータ-(tPA)のような生理活性物質の生産にとってはむしろDOが10mg/L以上の高いDOレベルの方が有利であることがわかった。さらに細胞増殖に適した溶存酸素濃度と生理活性物質の生産に適した溶存酸素濃度に分けて培養する効果についても検討し、効果があることがわかった。 4.EPOなどの生理活性物質を生産するための固定化動物細胞による高密度培養を行うにあたって、溶存酸素濃度を最適値に保つと共に、栄養源であるグルコースやアミノ酸の枯渇を防ぎ、阻害的な代謝産物である乳酸とアンモニア濃度を高めないように栄養源濃度の制御を行うことが重要となる。そこで我々が開発した多項目同時測定システムと溶存酸素制御システムとを用いて、栄養源濃度の制御の効果を調べた。その結果、ハイブリドーマ細胞の場合にはグルコースまたはグルタミン濃度を低く保つとモノクローナル抗体の生産性が向上することがわかった。

  24. 光で誘導可能な植物遺伝子発現システムの構築と有用物質生産への応用

    魚住 信之

    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: 名古屋大学

    1993 - 1993

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    光で誘導可能な植物遺伝子発現システムの構築として、申請者は以下の研究を行い、成果を得た。 1.トマト由来のribulose 1,5-bisphosphate carboxylase oxygenase/小サブユニット(rbsS)のプロモーターをbeta-グルクロニダーゼ(GUS)遺伝子上流に連結し、大腸菌及びAgrobacteriumで複製可能なpBI系のプラスミドに挿入した。構築したプラスミドをAgrobacterium tumefaciens及びA.rhizogenesを用いた遺伝子導入法で、タバコの細胞及びアジュガの植物体に感染させ、それぞれの遺伝子組換え体を得た。構築した遺伝子組換え体は明条件においてプラスミドに組み込まれたGUS遺伝子が発現し、その生産物であるbeta-グルクロニダーゼを生産することが確認された。 2.培養環境の最適化を目的とした計測法を確立するための初期検討として、知識工学を利用した不定胚分化状況の画像処理計測法を構築した。本システムは測定対象の画像からコンピュータによってパラメーターを抽出し、ニューラルネットワークによりその分化の度合いを決定する方法である。その結果、対象の分化の度合いをを効率よく決定することが可能であった。よって、本システムを培養中の細胞に適用することによって培養環境の監視が可能である示唆される。 3.1で作成した光誘導可能な遺伝子組換え体がバイオリアクターで有用蛋白質を大量生産することを目的とした高密度培養が可能であるか検討した。タバコ細胞を用いた検討ではバイオリアクターによる培養が可能であるものの撹拌翼の形状などなお検討を要することがわかった。

  25. Development of artificial seed using plant hairy root

    KOBAYASHI Takeshi, KIMURA Tatsuro, UOZUMI Nobuyuki

    Offer Organization: Japan Society for the Promotion of Science

    System: Grants-in-Aid for Scientific Research

    Category: Grant-in-Aid for Developmental Scientific Research (B)

    Institution: Nagoya University

    1991 - 1992

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    We focused on the development of micropropagation procedure of horseradish hairy root by using artificial seed system from the standpoint of bioengineering. 1. Auxin supplementation stimulated branching, which resulted in the increment of the number of apical meristem and cell growth rate. The kinetic growth model was developed to simulate the hairy root growth. 2. The root fragments with branch or apical meristem excised by blade were highly regenerated under dark conditions. 3 The adventitious shoot primordium on the horseradish hairy root by microscopic morphological observation and alteration to plantlet at the exposure to light is identified. The adventitious shoot primordium offers an advantage for relatively higher synchrony in development at exposure to light, compared with two other cells. It is necessary to increase the number of the adventitious shoot primordium formation in an artificial seed system. 4. Plantlet developed from hairy root were obtained by treatment cytokinin supplementation. To cut the root into fragments, the blender was used. The mechanical excision improved the bioprocess to produce the competent root fragments as artificial seed. the bioreactor, the regeneration frequency was similar to that of that in shake flask. The candidate hairy root cells included in beads were classified into three ; 1, the root fragment cultured under dark condition. 2, the adventitious shoot primordium formed under dark condition. 3, plantlet developed from the hairy root which was formed after transferred into light. The above results show the possibility of application of hairy root for artificial seed production.

  26. 遺伝子工学を応用した硝化脱窒過程の解析と高効率化

    飯島 信司, 魚住 信之, 小林 猛

    Offer Organization: 日本学術振興会

    System: 科学研究費助成事業

    Category: 重点領域研究

    Institution: 名古屋大学

    1991 - 1991

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    遺伝子工学を応用して窒素除去を好気的な硝化過程と嫌気的な脱窒過程を同時に行うことを目指して、亜硝酸還元酵素の遺伝子のクロ-ン化を試みている。前年度までにP.denitrificans由来の亜硝酸還元酵素を精製後、亜硝酸還元酵素遺伝子を含むDNA断片をクロ-ン化したが、N末端アミノ酸配列部位およびプロモ-タ-を含む5'ーflanking領域が欠失していた。本年度は、亜硝酸還元遺伝子全体をクロ-ン化して活性のある亜硝酸還元酵素を得るとともに、遺伝子があると期待される他の脱窒に必要な酵素遺伝子をクロ-ン化することを試みた。P.denitrificansの染色体DNAをXhoIで完全消化およびSa1Iで部分消化しλファ-ジを用いたXhoI、Sa1Iライブラリ-を作成し、クロ-ン化された遺伝子の隣接領域のDNAの分離を図った。 λファ-ジ上にクロ-ン化したP.denitrificans染色体のXhoIライブラリ-中から、pNO1上のSa1IーHind III 200bpのプロ-ブとハイブリダイズするクロ-ンλーH1、λーX2、またλファ-ジSa1Iライブラリ-から同様にハイブリダイズするクロ-ンλーS1、λーS2、λーS3、λーS4が得られた。一方、P.denitrificansの染色体DNAをXhoIおよびSa1Iで消化したパタ-ンよりこのプロ-ブに対応するXhoI断片は15kb、Sa1I断片は3kb程度と決定した。獲得した6種類のクロ-ンを、Sa1IおよびXhoIで完全消化してサザンハイブリダイゼイションを行ったところ、プロ-ブとハイブリダイズする5種類のXhoI断片およびSa1I断片のクロ-ン化DNAは目的DNAを保持していることを示していた。この5種類のクロ-ン化DNAの制限酵素パタ-ンを調べたところ、Hind IIIーSa1I断片以外が少しずつ異なっており、Hind IIIーSa1I断片を中心として前後に散らばっていることが明かとなり、既にクロ-ン化されている遺伝子の上流に最大3Kbの遺伝子が今回クロ-ン化された。

  27. セルロ-ス直接発酵微生物の育種とアルコ-ル燃料の高効率生産

    小林 猛, 魚住 信之, 飯島 信司

    Offer Organization: 日本学術振興会

    System: 科学研究費助成事業

    Category: 重点領域研究

    Institution: 名古屋大学

    1990 - 1990

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    1.本研究室でクロ-ン化した6種類のセルラ-ゼのうち、セロビオヒドロラ-ゼ遺伝子を酵母の分泌ベクタ-に連結したプラスミドを作成し酵母に導入した。本酵母が生産するセロビオヒドロラ-ゼの約40%が細胞外(培養液中)に分泌され、酵母で機能するプロモ-タ-や分泌シグナルを含まないセルラ-ゼ遺伝子を導入した組換え酵母に比べてセルラ-ゼ生産は約50倍に増大した。また、活性染色法からセロビオヒドロ-ラゼは細胞内外に存在し、分子量の異なる数種類のセルラ-ゼが生産されてした。特に培養上清に分泌された酵素のみに糖鎖が付加されていることが、糖鎖染色の結果から明らかとなった。 2.今回構築した組換か酵母は、酵母エキスとポリペプトンに糖源といてグルコ-スを添加した培地で最高の細胞増殖速度が達成され、同時に培養液中にセルラ-ゼ活性も最高の74U/lであった。将来実際にセルロ-スを資化する酵母を用いた場合にはグルコ-スの生成が律速となり、培養液中のグルコ-スは低濃度であると予想されるが、低濃度のグルコ-スにおいてもセルラ-ゼは生産された。更に本組換え酵母のセルロ-ス分解性を検討したところ、カルボキシメチルーセルロ-スを加えた培地においてセルラ-ゼ活性の増大とセルロ-スの分解に起因する培養液の粘度低下が観察され、セルロ-スの分解が起っていると推定された。 3.セルロ-ス資化酵母を使って解率的なアルコ-ル生産を行なうには、グルコ-ス濃度の制御が不可欠であることから、乳酸発酵においてグルコ-ス濃度制御を検討した。グルコ-ス・乳酸自動分析装置によりグルコ-スを一定濃度に保って乳酸菌の培養を行なったところ、グルコ-スが低濃度(1g/l)の場合には57g/lの乳酸が生成した。グルコ-スの定量は自動液クロ分析装置を用いた。一方、グルコ-スを高濃度(20g/l)に制御した場合は副産物であり増殖に阻害的な酢酸が蓄積し乳酸生産量も減少したので、グルコ-スを低濃度に制御することの重要性が明らかになった。

  28. 遺伝子工学を応用した硝化脱窒過程の解析と高効率化

    飯島 信司, 魚住 信之, 小林 猛

    Offer Organization: 日本学術振興会

    System: 科学研究費助成事業

    Category: 重点領域研究

    Institution: 名古屋大学

    1990 - 1990

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    1.前年度にクロ-ン化した亜硝酸還元酵素遺伝子を含むと考えられる11.5kbp <Hind>___ーIII フラグメントをサブクロ-ン化し、詳細な制限酵素地図を作製した。クロ-ニングに使用したオリゴヌクレオチドとクロ-ン化したDNAフラグメントの相同性をサザンハイブリダイゼ-ション法で検討した。この結果オリゴヌクレオチドは、クロ-ン化DNAの端200bpと相同性があることが判明した。そこでこのフラグメント周辺のDNA塩基配列をダイデオキシ法で決定した結果、DNA挿入断片の端から130bp付近にプロ-ブと相補的な塩基配列が存在することがわかった。また塩基配列から推定されるこの前後のアミノ酸配列はさきに精製した酵素のアミノ酸配列と一致し、クロ-ン化されたDNAフラグメントは亜硝酸還元酵素遺伝子に相違ないことが確認された。 2.クロ-ン化した亜硝酸還元酵素遺伝子を<trp>___ーおよび<lacZ>___ープロモ-タ-を含む発現ベクタ-に連結し大腸菌に導入した後、ウエスタンブロッティングにより目的遺伝子産物の発現を検出した。その結果、亜硝酸還元酵素抗原のみかけの分子量は約5万であった。活性型亜硝酸還元酵素の分子量は7万であるので、クロ-ン化したDNAフラグメントは酵素全体の約2/3を含んでいると考えられる。また決定したDNA塩基配列から推定されるアミノ酸配列からこの亜硝酸還元酵素抗原は活性型のN末端側が欠如していると考えられ、本酵素の構造遺伝子は、11.5kb <Hind>___ーIIIフラグメント内の4kbp程度の領域に含まれていると推定された。上記実験全般にわたりプラスミドDNAの大量調整はカゴ型攪拌リアクタ-(培養槽)を使用した。

  29. Development of Suitable Bioreactors for Plant Hairy Roots and Production of Useful Biochemicals.

    KOBAYASHI Takeshi, HONDA Hiroyuki, UOZUMI Nobuyuki

    Offer Organization: Japan Society for the Promotion of Science

    System: Grants-in-Aid for Scientific Research

    Category: Grant-in-Aid for General Scientific Research (B)

    Institution: Nagoya University

    1989 - 1990

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    Plant hairy root, transgenic plant induced by Agrobacterium rhizogenes, have become of interest as a candidate for in vitro culture plant or organs. Hairy root cultivation is required for a high oxygen transfer coefficient and low shear stress. To meet these requirements, we have developed a turbine blade reactor and an immobilized rotating drum reactor. Using these bioreactors, we have Obtained high cell density culture in carrot hairy roots, approximately 2.5 fold compared with culture in a shake flask. We have induced hairy roots from various plant cells to find a high content of useful enzyme, peroxidase, in Pak-bung hairy root. Light stimulated peroxidase production, and NaCl addition to medium enhanced the peroxidase excretion. The total peroxidase production could be greatly improved by adsorption of peroxidase in horseradish hairy roots culture medium with hydrophobic resin in these combined condition.

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    耐塩性植物の構築