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Physiological functions and selective substrate-cleavage mechanism of E. coli intramembrane protease RseP

Research Project

Project/Area Number 18H02404
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Review Section Basic Section 43030:Functional biochemistry-related
Research InstitutionKyoto University

Principal Investigator

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

Co-Investigator(Kenkyū-buntansha) 檜作 洋平  京都大学, ウイルス・再生医科学研究所, 助教 (70568930)
Project Period (FY) 2018-04-01 – 2021-03-31
Project Status Completed (Fiscal Year 2020)
Budget Amount *help
¥17,420,000 (Direct Cost: ¥13,400,000、Indirect Cost: ¥4,020,000)
Fiscal Year 2020: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000)
Fiscal Year 2019: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000)
Fiscal Year 2018: ¥6,760,000 (Direct Cost: ¥5,200,000、Indirect Cost: ¥1,560,000)
Keywords膜内切断プロテアーゼ / 大腸菌 / 切断基質 / 膜内部でのタンパク質切断 / S2Pプロテアーゼ / Fecオペロン / 鉄取り込み / 蛍光ペプチド / 速度論的解析 / 小分子膜タンパク質 / RseP / Toxin-antitoxin / HokB / Persister / small membrane protein / FRET / 生体膜 / プロテアーゼ / 膜内切断 / 蛍光基質
Outline of Final Research Achievements

Intramembrane cleaving proteases (I-CLiPs) are known to act in a variety of important cellular processes in diverse organisms. RseP, an Escherichia coli S2P family intramembrane cleaving protease, is involved in the regulation of the sigma E pathway extracytoplasmic stress response and membrane quality control through specific cleavage of substrates within the lipid bilayer. We have identified novel substrates of RseP and analyzed the physiological significance of their RseP-dependent cleavage. Also, we tried to construct an in vitro assay system that enables quantitative analysis of the RseP activity. These results would contribute to the elucidation of the cellular roles of and the mechanism underlying proteolysis by RseP.

Academic Significance and Societal Importance of the Research Achievements

本研究は、申請者ら独自の発見・研究成果を直接踏まえ展開するもので、BepA機能解明を通じて、大腸菌等グラム陰性細菌の外膜形成・維持機構のみならず、protease制御機構に新たな知見を供し、基礎から応用まで広くインパクトを与えるだろう。本研究は、新奇発見に基づくアプローチによる独創性と、独自の成果を統合・発展させる着実性の、両面を兼ね備えたものである。

Report

(4 results)
  • 2020 Annual Research Report   Final Research Report ( PDF )
  • 2019 Annual Research Report
  • 2018 Annual Research Report
  • Research Products

    (24 results)

All 2021 2020 2019 2018

All Journal Article (12 results) (of which Peer Reviewed: 11 results,  Open Access: 6 results) Presentation (12 results) (of which Int'l Joint Research: 1 results)

  • [Journal Article] Moving toward generalizable NZ-1 labeling for 3D structure determination with optimized epitope tag insertion.2021

    • Author(s)
      Tamura-Sakaguchi, R., Aruga, R., Hirose, M., Ekimoto, T., Miyake, T., Hizukuri, Y., Oi, R., Kaneko, M. M., Kato, Y., Akiyama, Y., Ikeguchi, M., Iwasaki, K., and Nogi, T.
    • Journal Title

      Acta Crystallogr. D

      Volume: -

    • NAID

      120007032351

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Escherichia coli S2P family intramembrane protease RseP is engaged in the regulated sequential cleavages of FecR in the ferric citrate signaling.2021

    • Author(s)
      Yokoyama, T., Niinae, T., Tsumagari, K., Imami, K., Ishihama, Y., Hizukuri, Y., and Akiyama, Y.
    • Journal Title

      J. Biol. Chem.

      Volume: -

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Reversible auto-inhibitory regulation of Escherichia coli metallopeptidase BepA for selective β-barrel protein degradation.2020

    • Author(s)
      Daimon 他7名
    • Journal Title

      Proc. Natl. Acad. Sci. USA.

      Volume: 117 Issue: 45 Pages: 27989-27996

    • DOI

      10.1073/pnas.2010301117

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Involvement of a Membrane-Bound Amphiphilic Helix in Substrate Discrimination and Binding by an Escherichia coli S2P Peptidase RseP2020

    • Author(s)
      Miyake Takuya, Hizukuri Yohei, Akiyama Yoshinori
    • Journal Title

      Frontiers in Microbiology

      Volume: 11 Pages: 607381-607381

    • DOI

      10.3389/fmicb.2020.607381

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Fine interaction profiling of VemP and mechanisms responsible for its translocation-coupled arrest-cancelation2020

    • Author(s)
      Miyazaki Ryoji、Akiyama Yoshinori、Mori Hiroyuki
    • Journal Title

      eLife

      Volume: 9

    • DOI

      10.7554/elife.62623

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] 大腸菌表層ストレス応答:インプットとアウトプットに働くプロテアーゼの機能2020

    • Author(s)
      秋山芳展
    • Journal Title

      IFO Res. Commun.

      Volume: 33 Pages: 217-220

    • Related Report
      2019 Annual Research Report
  • [Journal Article] An in vivo protease activity assay for investigating the functions of the Escherichia coli membrane protease HtpX2019

    • Author(s)
      Yoshitani Kohei, Hizukuri Yohei, Akiyama Yoshinori
    • Journal Title

      FEBS Letters

      Volume: 593 Issue: 8 Pages: 842-851

    • DOI

      10.1002/1873-3468.13368

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Structural basis of the function of the β-barrel assembly-enhancing protease BepA2019

    • Author(s)
      Mohammad Shahrizal, Yasushi Daimon, Yoshiki Tanaka, Yugo Hayashi, Shintaro Nakayama, Shigehiro Iwaki, Shin-ichiro Narita, Hironari Kamikubo, Yoshinori Akiyama, Tomoya Tsukazaki
    • Journal Title

      Journal of Molecular Biology

      Volume: 431 Issue: 3 Pages: 625-635

    • DOI

      10.1016/j.jmb.2018.11.024

    • Related Report
      2019 Annual Research Report 2018 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Heat shock transcription factor σ<sup>32</sup> defective in membrane transport can be suppressed by transposon insertion into genes encoding a restriction enzyme subunit or a putative autotransporter in <i>Escherichia coli</i>2018

    • Author(s)
      Yura Takashi、Miyazaki Ryoji、Fujiwara Keigo、Ito Koreaki、Chiba Shinobu、Mori Hiroyuki、Akiyama Yoshinori
    • Journal Title

      Genes & Genetic Systems

      Volume: 93 Issue: 6 Pages: 229-235

    • DOI

      10.1266/ggs.18-00040

    • NAID

      130007582071

    • ISSN
      1341-7568, 1880-5779
    • Year and Date
      2018-12-01
    • Related Report
      2018 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Identification of an Internal Cavity in the PhoQ Sensor Domain for PhoQ Activity and SafA-mediated Control2018

    • Author(s)
      K. Yoshitani, E. Ishii, K. Taniguchi, H. Sugimoto, Y. Shiro, H. Mori, Y. Akiyama, A. Kato, R. Utsumi, Y. Eguchi
    • Journal Title

      Bioscience, Biotechnology, and Biochemistry

      Volume: 83 Issue: 4 Pages: 684-694

    • DOI

      10.1080/09168451.2018.1562879

    • Related Report
      2019 Annual Research Report 2018 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Identification and characterization of arrest motif in VemP by systematic mutational analysis.2018

    • Author(s)
      Mori, H., Sakashita, S., Ito, J., Ishii, E. and Akiyama, Y.
    • Journal Title

      J. Biol. Chem.

      Volume: 293 Issue: 8 Pages: 2915-2926

    • DOI

      10.1074/jbc.m117.816561

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed
  • [Journal Article] A new photo-cross-linking approach for analysis of protein dynamics in vivo.2018

    • Author(s)
      Miyazaki, R., Myogo, N., Mori, H. and Akiyama, Y.
    • Journal Title

      J. Biol. Chem.

      Volume: 293 Issue: 2 Pages: 677-686

    • DOI

      10.1074/jbc.m117.817270

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed
  • [Presentation] 新生ポリペプチド鎖依存的な膜局在化によるmRNA分解促進2020

    • Author(s)
      石井英治、秋山芳展、森 博幸
    • Organizer
      日本農芸化学会2020年度大会
    • Related Report
      2020 Annual Research Report
  • [Presentation] 菌の外膜生合成と品質管理に関わる2機能性プロテアーゼBepAの基質認識機構2020

    • Author(s)
      宮﨑亮次、吉谷亘平、森 博幸、秋山芳展
    • Organizer
      第93回日本生化学会大会
    • Related Report
      2020 Annual Research Report
  • [Presentation] 大腸菌膜内切断プロテアーゼRsePを介したTA systemの制御: 細胞休眠・覚醒における意義2020

    • Author(s)
      檜作洋平、横山 達彦、秋山芳展
    • Organizer
      第93回日本生化学会大会
    • Related Report
      2020 Annual Research Report
  • [Presentation] 大腸菌S2PファミリープロテアーゼRsePの基質探索:鉄取り込みに関与する転写制御因子の切断とその生理的意義2020

    • Author(s)
      横山達彦、新苗智也、津曲和哉、今見考志、石濱泰、檜作洋平、秋山芳展
    • Organizer
      第93回日本生化学会大会
    • Related Report
      2020 Annual Research Report
  • [Presentation] 大腸菌膜内切断プロテアーゼRsePの基質切断と構造維持に重要な膜表在・両親媒性ヘリックスPCT-H1の解析2019

    • Author(s)
      7.三宅拓也、檜作洋平、秋山芳展
    • Organizer
      第16回21世紀大腸菌研究会
    • Related Report
      2019 Annual Research Report
  • [Presentation] FRET蛍光ペプチドを用いた細菌S2Pファミリー膜内切断プロテアーゼのリアルタイム切断kinetics計測系の構築2019

    • Author(s)
      10.檜作洋平、秋山芳展
    • Organizer
      第92回日本生化学会大会
    • Related Report
      2019 Annual Research Report
  • [Presentation] 大腸菌膜内切断プロテアーゼRsePの基質切断と構造維持に重要な膜表在・両親媒性ヘリックスの解析2019

    • Author(s)
      12.三宅拓也、檜作洋平、秋山芳展
    • Organizer
      第92回日本生化学会大会
    • Related Report
      2019 Annual Research Report
  • [Presentation] 大腸菌膜内切断プロテアーゼRsePの新規基質切断とその生理的意義: 低分子膜タンパク質HokBの切断を介したパーシスター形成制御2019

    • Author(s)
      13.横山達彦、檜作洋平、秋山芳展
    • Organizer
      第92回日本生化学会大会
    • Related Report
      2019 Annual Research Report
  • [Presentation] Kinetic analysis of the proteolytic reaction catalyzed by S2P family intramembrane protease RseP using a FRET-based real-time assay system2019

    • Author(s)
      14.檜作洋平、秋山芳展
    • Organizer
      第57回日本生物物理学会年会
    • Related Report
      2019 Annual Research Report
  • [Presentation] 大腸菌膜内切断プロテアーゼRsePの新規機能調節領域の解析2018

    • Author(s)
      三宅拓也、檜作洋平、秋山芳展
    • Organizer
      第15回21世紀大腸菌研究会
    • Related Report
      2018 Annual Research Report
  • [Presentation] Functional and structural roles of a membrane-periplasm interface domain in RseP, an Escherichia coli S2P family intramembrane protease.2018

    • Author(s)
      Miyake, T., Hizukuri, Y. and Akiyama, Y.
    • Organizer
      International symposium Proteins: From the Cradle to the Grave
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research
  • [Presentation] 大腸菌膜内切断プロテアーゼRsePの構造維持と機能調節に関わる新規ドメインの解析2018

    • Author(s)
      三宅拓也、檜作洋平、秋山芳展
    • Organizer
      第91回日本生化学会大会
    • Related Report
      2018 Annual Research Report

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Published: 2018-04-23   Modified: 2022-01-27  

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