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Live-cell imaging of a bacterial extracytoplasmic stress response and exploration of a novel function of an intramembrane protease that regulates the stress response

Research Project

Project/Area Number 26840033
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeMulti-year Fund
Research Field Functional biochemistry
Research InstitutionKyoto University

Principal Investigator

HIZUKURI Yohei  京都大学, ウイルス・再生医科学研究所, 助教 (70568930)

Project Period (FY) 2014-04-01 – 2017-03-31
Project Status Completed (Fiscal Year 2016)
Budget Amount *help
¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2016: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2015: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2014: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Keywords大腸菌 / プロテアーゼ / 表層ストレス応答 / S2P / 蛍光イメージング / GFP / 生化学 / 生物物理学 / small membrane protein / 膜内切断プロテアーゼ / サルモネラ菌 / 酸性ストレス応答 / 一細胞リアルタイム観察
Outline of Final Research Achievements

An intramembrane protease RseP is a key regulator in the σE-pathway extracytoplasmic stress response (ESR) which is critical for bacterial survival under stressed conditions. In this research project, I have aimed to understand a whole picture of the ESR system. Through a detailed functional analysis of the Escherichia coli RseP, I have revealed that two novel regions in RseP, the MRE β-loop and the C1N regions, play a crucial role for substrate recognition and cleavage mechanism. This finding gives us new insight into intramembrane cleaving proteases highly conserved from bacteria to human. We have identified some novel proteolytic substrates of RseP, which might taught us possible new roles of RseP. I have also established a live-cell imaging system for visualizing an RseP-dependent substrate cleavage event by fluorescent microscopy. This system must be useful for a kinetic analysis for the ESR system in the future.

Report

(4 results)
  • 2016 Annual Research Report   Final Research Report ( PDF )
  • 2015 Research-status Report
  • 2014 Research-status Report
  • Research Products

    (17 results)

All 2017 2016 2015 2014 Other

All Journal Article (2 results) (of which Peer Reviewed: 2 results,  Acknowledgement Compliant: 2 results,  Open Access: 1 results) Presentation (8 results) Book (2 results) Remarks (5 results)

  • [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] 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 Research-status Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Presentation] Screening of physiological substrates of E. coli rhomboid protease GlpG: possible involvement of GlpG in the flagellar function2016

    • Author(s)
      Yohei Hizukuri, Kosuke Terushima, Yoshinori Akiyama
    • Organizer
      第54回日本生物物理学会年会
    • Place of Presentation
      つくば国際会議場(茨城県)
    • Year and Date
      2016-11-25
    • Related Report
      2016 Annual Research Report
  • [Presentation] 細菌膜内切断プロテアーゼの総合的理解に向けて: ~機能制御機構を中心に~2016

    • Author(s)
      檜作 洋平
    • Organizer
      ウイルス感染症・生命科学先端融合的共同研究拠点セミナー
    • Place of Presentation
      京都大学 ウイルス・再生医科学研究所(京都)
    • Year and Date
      2016-11-24
    • Related Report
      2016 Annual Research Report
  • [Presentation] 細菌RhomboidプロテアーゼGlpGの生理的切断基質の探索及びべん毛III型分泌装置との関係2016

    • Author(s)
      檜作 洋平、照島 功祐、秋山 芳展
    • Organizer
      第89回日本生化学会大会
    • Place of Presentation
      仙台国際センター/東北大学川内北キャンパス(宮城)
    • Year and Date
      2016-09-25
    • Related Report
      2016 Annual Research Report
  • [Presentation] 大腸菌RhomboidプロテアーゼGlpGの生理的切断基質の探索及び基質候補の解析2016

    • Author(s)
      檜作 洋平、照島 功祐、秋山 芳展
    • Organizer
      21世紀大腸菌研究会
    • Place of Presentation
      グリーンピア南阿蘇(熊本)
    • Year and Date
      2016-06-02
    • Related Report
      2016 Annual Research Report
  • [Presentation] 細菌表層ストレス応答を制御するS2Pファミリー膜内切断プロテアーゼの細胞内イメージング解析2015

    • Author(s)
      檜作 洋平,秋山 芳展
    • Organizer
      BMB2015
    • Place of Presentation
      神戸ポートアイランド(兵庫)
    • Year and Date
      2015-12-01
    • Related Report
      2015 Research-status Report
  • [Presentation] Live-cell imaging of a proteolytic event by an intramembrane protease RseP that regulates extracytoplasmic stress response2015

    • Author(s)
      檜作 洋平,秋山 芳展
    • Organizer
      第53回日本生物物理学会年会
    • Place of Presentation
      金沢大学 角間キャンパス 自然科学本館(石川)
    • Year and Date
      2015-09-13
    • Related Report
      2015 Research-status Report
  • [Presentation] 表層ストレス応答制御に関与するRsePプロテアーゼの基質切断反応の細胞内蛍光イメージングに向けた取り組み2015

    • Author(s)
      檜作 洋平,秋山 芳展
    • Organizer
      第12回 21世紀大腸菌研究会
    • Place of Presentation
      琵琶湖グランドホテル・京近江(滋賀)
    • Year and Date
      2015-06-04
    • Related Report
      2015 Research-status Report
  • [Presentation] 細菌表層ストレス応答制御に関わる膜内切断プロテアーゼRsePのタンデムPDZドメインによる切断基質選別機構の解析2014

    • Author(s)
      檜作洋平,小田隆,田畑早苗,川上-田村恵子,大井里香,佐藤衛,高木淳一,禾晃和,秋山芳展
    • Organizer
      第87回日本生化学会大会
    • Place of Presentation
      京都,国立京都国際会館・グランドプリンスホテル京都
    • Year and Date
      2014-10-15 – 2014-10-18
    • Related Report
      2014 Research-status Report
  • [Book] Methods in Enzymology, Enzymology at the Membrane Interface: Intramembrane Proteases (Edited by Michael H. Gelb)内, Chapter One - Biochemical Characterization of Function and Structure of RseP, an Escherichia coli S2P Protease.2017

    • Author(s)
      Yohei Hizukuri, Koichiro Akiyama, Yoshinori Akiyama
    • Total Pages
      496
    • Publisher
      Elsevier Inc.
    • Related Report
      2016 Annual Research Report
  • [Book] 別冊・医学のあゆみ「レドックスUPDATE ―ストレス制御の臨床医学・健康医学」(監修:平家 俊男,淀井 淳司)内,「膜内部での蛋白質分解とストレス応答制御」の項2015

    • Author(s)
      檜作 洋平,秋山 芳展
    • Total Pages
      344
    • Publisher
      医歯薬出版株式会社
    • Related Report
      2015 Research-status Report
  • [Remarks] 京都大学 ウイルス・再生医科学研究所 生体膜システム分野 秋山研究室ホームページ

    • URL

      http://www.virus.kyoto-u.ac.jp/Lab/akiyama/

    • Related Report
      2016 Annual Research Report
  • [Remarks] 京都大学 ウイルス・再生医科学研究所 生体膜システム分野 紹介ページ

    • URL

      http://www.infront.kyoto-u.ac.jp/research/lab31/

    • Related Report
      2016 Annual Research Report
  • [Remarks] 京都大学大学院理学研究科化学専攻 生体分子動態化学分科紹介

    • URL

      http://www.kuchem.kyoto-u.ac.jp/organization/lab/virus.html

    • Related Report
      2016 Annual Research Report
  • [Remarks] 京都大学ウイルス研究所秋山ラボHP

    • URL

      http://www.virus.kyoto-u.ac.jp/Lab/akiyama/index.html

    • Related Report
      2015 Research-status Report
  • [Remarks] 京都大学ウイルス研究所秋山研究室ホームページ

    • URL

      http://www.virus.kyoto-u.ac.jp/Lab/akiyama/index.html

    • Related Report
      2014 Research-status Report

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Published: 2014-04-04   Modified: 2018-03-22  

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