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ビブリオ菌における新生鎖機能を介したタンパク質膜透過の制御

Publicly Offered Research

Project AreaNascent-chain Biology
Project/Area Number 15H01532
Research Category

Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)

Allocation TypeSingle-year Grants
Review Section Biological Sciences
Research InstitutionKyoto University

Principal Investigator

秋山 芳展  京都大学, ウイルス・再生医科学研究所, 教授 (10192460)

Project Period (FY) 2015-04-01 – 2017-03-31
Project Status Completed (Fiscal Year 2016)
Budget Amount *help
¥14,170,000 (Direct Cost: ¥10,900,000、Indirect Cost: ¥3,270,000)
Fiscal Year 2016: ¥7,410,000 (Direct Cost: ¥5,700,000、Indirect Cost: ¥1,710,000)
Fiscal Year 2015: ¥6,760,000 (Direct Cost: ¥5,200,000、Indirect Cost: ¥1,560,000)
Keywords翻訳停止 / リボソーム / 新生鎖 / ビブリオ菌 / SecDF / タンパク質 / 翻訳 / 細菌 / VemP
Outline of Annual Research Achievements

VemP (Vibrio protein export monitoring polypeptide)は、159アミノ酸残基からなる分泌タンパク質であり、N末端にシグナル配列を、C末端にビブリオ属細菌間で高度に保存された領域を持つ。VemPは、ビブリオ菌のタンパク質膜透過能を自身の膜透過能としてモニターし、膜透過不全時には、C末端近傍で自身の翻訳を安定に低下させ、下流のsecDF2遺伝子の発現を上昇する役割を持つ。VemPの翻訳停止は、C末端保存領域とリボソームトンネル内壁との特異的な相互作用により生じ、この相互作用にはC末端保存領域内の多数のアミノ酸残基の関与が示唆されていたが、詳細な分子機構については未だ不明であった。
本年度は、VemP C末端保存領域の翻訳停止における役割を明らかにするために、翻訳停止の状態を酵素活性として評価できるdelta ss-vemP'-lacZレポーター系を構築し、実験系の有用性を確認した。このシステムを用いて、C末端保存領域20アミノ酸残基を対象とした網羅的・系統的な変異解析を行い、各アミノ酸残基の翻訳停止における役割を検討した。その結果、ほぼ全ての置換を容認しない極めて重要なアミノ酸残基の存在を明らかにすると共に、この領域が二次構造を形成している可能性を強く示唆する結果を得た。この領域が仮にαヘリックス構造を形成していると仮定すると、このC末端保存領域は、強い両親媒性の特徴を持ち、翻訳停止に必須のアミノ酸残基は疎水性面に集中しているという明確な特徴が見られた。

Research Progress Status

28年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

28年度が最終年度であるため、記入しない。

Report

(2 results)
  • 2016 Annual Research Report
  • 2015 Annual Research Report
  • Research Products

    (9 results)

All 2017 2016 2015

All Journal Article (5 results) (of which Peer Reviewed: 5 results,  Open Access: 2 results,  Acknowledgement Compliant: 4 results) Presentation (4 results) (of which Int'l Joint Research: 2 results,  Invited: 4 results)

  • [Journal Article] Biochemical Characterization of Function and Structure of RseP, an Escherichia coli S2P Protease2017

    • Author(s)
      Hizukuri, Y. , Akiyama, K. and Akiyama, Y.
    • Journal Title

      Methods in Enzymology

      Volume: 584 Pages: 1-33

    • DOI

      10.1016/bs.mie.2016.09.044

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] Involvement of a conserved GFG motif region in substrate binding by RseP, an Escherichia coli S2P protease2017

    • Author(s)
      Akiyama, K., Hiuzkuri, Y. and Akiyama, Y.
    • Journal Title

      Molecular Microbiology

      Volume: 印刷中 Issue: 5 Pages: 737-751

    • DOI

      10.1111/mmi.13659

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] A novel SRP recognition sequence in the homeostatic control region od heat shock transcription factor sigma 322016

    • Author(s)
      Miyazaki, R., Yura, T., Suzuki, T. Dohmae, N., Mori, H. and Akiyama, Y.
    • Journal Title

      Scientfic reports

      Volume: 6 Issue: 1 Pages: 24147-24147

    • DOI

      10.1038/srep24147

    • Related Report
      2016 Annual Research Report 2015 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Nascent chain-monitored remodeling of the Sec machinery for salinity adaptation of marine bacteria2015

    • Author(s)
      Ishii, E., Chiba, S., Hashimoto, N., Kojima, S., Homma, M., Ito, K., Akiyama, Y. and Mori, H.
    • Journal Title

      Proc. Natl. Acad. Sci. USA

      Volume: 112 Issue: 40

    • DOI

      10.1073/pnas.1513001112

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] Roles of the membrane-reentrant β-hairpin-like loop of RseP protease in selective substrate cleavage2015

    • Author(s)
      Koichiro Akiyama, Shinya Mizuno, Yohei Hizukuri, Hiroyuki Mori, Terukazu Nogi, Yoshinori Akiyama
    • Journal Title

      eLife

      Volume: 4

    • DOI

      10.7554/elife.08928

    • NAID

      120005712942

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Presentation] Regulation of E. coli heat shock response regulator sigma 32 via SRP protein targeting pathway2016

    • Author(s)
      Akiyama, Y.
    • Organizer
      23rd East Asia Joint Symposium / 15th Cross-Strait Symposium on Biochemical Research / 13th Symposium of the Frontiers of Biomedical Sciences
    • Place of Presentation
      Taipei
    • Year and Date
      2016-10-18
    • Related Report
      2016 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Nascent chain-monitored remodeling of the Sec machinery for salinity adaptation of marine bacteria.2016

    • Author(s)
      Mori, H., Ishii, E., Chiba, S., Sakashita, S., Ito, K. & Akiyama, Y.
    • Organizer
      NASCENT CHAIN BIOLOGY
    • Place of Presentation
      山梨
    • Year and Date
      2016-09-01
    • Related Report
      2016 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Environmental adaptation of marine Vibrio by nascent chain-mediated remodeling of protein export machinery.2016

    • Author(s)
      Mori, H.
    • Organizer
      RNA2016 satellite-symposium “Nascent biology and Ribosome functions”
    • Place of Presentation
      京都
    • Related Report
      2016 Annual Research Report
    • Invited
  • [Presentation] Nascent chain-monitored remodeling of the Sec machinery for salinity adaptation of marine bacteria.2015

    • Author(s)
      Mori, H., Ishii, E., Chiba, S., Hashimoto, N., Kojima, S., Homma, M., Ito, K. and Akiyama, Y.
    • Organizer
      BMB2015 (第38回日本分子生物学会年会、第88回日本生化学会大会 合同大会) Symposium “Nascent chains: the ribosome as a hub for protein quality control”
    • Place of Presentation
      神戸
    • Year and Date
      2015-12-01
    • Related Report
      2015 Annual Research Report
    • Invited

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Published: 2015-04-16   Modified: 2018-03-28  

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