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Structural and functional analyses of a core complex of the bacterial flagellar export gate complex

Research Project

Project/Area Number 18K14638
Research Category

Grant-in-Aid for Early-Career Scientists

Allocation TypeMulti-year Fund
Review Section Basic Section 43020:Structural biochemistry-related
Research InstitutionOsaka University

Principal Investigator

Kinoshita Miki  大阪大学, 生命機能研究科, 特任研究員(常勤) (30790985)

Project Period (FY) 2018-04-01 – 2020-03-31
Project Status Completed (Fiscal Year 2019)
Budget Amount *help
¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2019: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Fiscal Year 2018: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Keywords細菌 / 蛋白質 / 遺伝学 / 電子顕微鏡 / 感染症
Outline of Final Research Achievements

The bacterial flagellar export gate complex is composed of five membrane proteins, FlhA, FlhB, FliP, FliQ and FliR, and utilizes proton motive force across the cytoplasmic membrane as the energy source to drive flagellar protein export. In this study, I investigated how the protein export channel of the export gate complex is opened to allow export substrates to cross the cytoplasmic membrane. I have provided experimental evidence suggesting that the C-terminal cytoplasmic domains of FlhA and FlhB not only mediates a hierarchical protein targeting but also regulate gate opening of the FliPQR protein export channel complex.

Academic Significance and Societal Importance of the Research Achievements

腸管出血性病原性大腸菌O157などの病原細菌による感染症は社会問題の一つで、それらの感染機構の解明ならびに克服のための方法の探索が続けられている。べん毛タンパク質輸送装置と高い相同性を示すIII型分泌装置が急性胃腸炎の発症に深く関わっている。本研究成果に基づいて病原性細菌のIII型分泌装置のゲート開閉機構を特異的に不活性化できれば、病原性細菌の病原性のみを破壊できる。したがって、従来の抗生物質のように細菌を死滅させることがないため耐性菌が発生しづらく、感染症の予防を含めた、これまでにない新しい治療法の開発につながると期待できる。

Report

(3 results)
  • 2019 Annual Research Report   Final Research Report ( PDF )
  • 2018 Research-status Report
  • Research Products

    (21 results)

All 2020 2019 2018 Other

All Int'l Joint Research (2 results) Journal Article (7 results) (of which Peer Reviewed: 7 results,  Open Access: 7 results) Presentation (10 results) (of which Int'l Joint Research: 2 results) Remarks (2 results)

  • [Int'l Joint Research] ユタ大学(米国)

    • Related Report
      2019 Annual Research Report
  • [Int'l Joint Research] 南京農業大学(中国)

    • Related Report
      2019 Annual Research Report
  • [Journal Article] FliK-Driven Conformational Rearrangements of FlhA and FlhB Are Required for Export Switching of the Flagellar Protein Export Apparatus2020

    • Author(s)
      Minamino Tohru、Inoue Yumi、Kinoshita Miki、Namba Keiichi
    • Journal Title

      Journal of Bacteriology

      Volume: 202 Issue: 3

    • DOI

      10.1128/jb.00637-19

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] The flexible linker of the secreted FliK ruler is required for export switching of the flagellar protein export apparatus.2020

    • Author(s)
      Kinoshita M, Tanaka S, Inoue Y, Namba K, Aizawa SI, Minamino T.
    • Journal Title

      Scientific Reports

      Volume: 10 Issue: 1 Pages: 838-838

    • DOI

      10.1038/s41598-020-57782-5

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Structural insights into the substrate specificity switch mechanism of the type III protein export apparatus2019

    • Author(s)
      Y., Y. Ogawa, M. Kinoshita, N. Terahara, M. Shimada, N. Kodera, T. Ando, K. Namba, A. Kitao, K. Imada, T. Minamino
    • Journal Title

      Structure

      Volume: 印刷中 Issue: 6 Pages: 965-976

    • DOI

      10.1016/j.str.2019.03.017

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Mutational analysis of the C‐terminal cytoplasmic domain of FlhB, a transmembrane component of the flagellar type III protein export apparatus in Salmonella2019

    • Author(s)
      Inoue Yumi、Kinoshita Miki、Namba Keiichi、Minamino Tohru
    • Journal Title

      Genes to Cells

      Volume: 24 Issue: 6 Pages: 408-421

    • DOI

      10.1111/gtc.12684

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Directional Switching Mechanism of the Bacterial Flagellar Motor2019

    • Author(s)
      Minamino Tohru、Kinoshita Miki、Namba Keiichi
    • Journal Title

      Computational and Structural Biotechnology Journal

      Volume: 17 Pages: 1075-1081

    • DOI

      10.1016/j.csbj.2019.07.020

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Structural insight into the substrate specificity switching mechanism of the type III protein export apparatus2019

    • Author(s)
      Inoue Y, Ogawa Y, Kinoshita M, Terahara N, Shimada M, Kodera N, Ando T, Namba K, Kitao A, Imada K, Minamino T
    • Journal Title

      Structure

      Volume: 印刷中

    • Related Report
      2018 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Structure and function of a bacterial flagellar export chaperone FlgN2018

    • Author(s)
      南野徹, 木下実紀
    • Journal Title

      生化学

      Volume: 90 Issue: 2 Pages: 203-206

    • DOI

      10.14952/SEIKAGAKU.2018.900203

    • NAID

      40021617516

    • ISSN
      0037-1017
    • Year and Date
      2018-04-25
    • Related Report
      2018 Research-status Report
    • Peer Reviewed / Open Access
  • [Presentation] Structural and functional analyses of the flagellar transmembrane export gate complex of the Salmonella flagellar type III protein export apparatus.2019

    • Author(s)
      Kinoshita M, Miyata T, Kawamoto A, Kato T, Namba K, Minamino T.
    • Organizer
      Gordon Research Conference on Salmonella Biology and Pathogenesis.
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Autocatalytic cleavage of FlhB is required for conformational rearrangements of FlhA to switch export specificity of the flagellar protein export apparatus.2019

    • Author(s)
      Minamino T, Inoue Y, Kinoshita M, Namba K.
    • Organizer
      Gordon Research Conference on Salmonella Biology and Pathogenesis.
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Gating mechanism of the bacterial flagellar protein export apparatus (バクテリアべん毛タンパク質輸送装置のゲート機構).2019

    • Author(s)
      Kinoshita M, Namba K, Minamino T.
    • Organizer
      第57回日本生物物理学会年会
    • Related Report
      2019 Annual Research Report
  • [Presentation] バクテリアべん毛タンパク質輸送装置のエネルギー共役機構.2019

    • Author(s)
      南野徹, 木下実紀, 難波啓一
    • Organizer
      日本生体エネルギー研究会第45回討論会
    • Related Report
      2019 Annual Research Report
  • [Presentation] クライオ電子顕 微鏡で解き明かす細菌べん毛モーター回転子の立体構造と回転対象性(Structure and rotational symmetry of the rotor of the bacterial flagellar motor revealed by electron cryomicroscopy)2018

    • Author(s)
      川本晃大, 宮田知子, 木下実紀, 南野徹, 加藤貴之, 難波啓一
    • Organizer
      日本顕微鏡学会第74 回学術講演会
    • Related Report
      2018 Research-status Report
  • [Presentation] べん毛先端のキャップ構造(The cap structure of the bacterial flagellar filament)2018

    • Author(s)
      牧野文信, 木田葵, 木下実紀, 宮田知子, 加藤貴之, 南野徹, 難波啓一, 今田勝巳
    • Organizer
      第18回日本蛋白質科学会年会
    • Related Report
      2018 Research-status Report
  • [Presentation] クライオ電子顕微鏡で解き明かす細菌べん毛モーター回転子の立体構造と回転対象性(Structure and rotational symmetry of the rotor of the bacterial flagellar motor revealed by electron cryomicroscopy)2018

    • Author(s)
      川本晃大, 宮田知子, 木下実紀, 南野徹, 加藤貴之, 難波啓一
    • Organizer
      第18回日本蛋白質科学会年会
    • Related Report
      2018 Research-status Report
  • [Presentation] Structural and functional analyses of the bacterial flagellar type III export gate complex (バクテリアべん毛輸送ゲート複合体の構造機能解析)2018

    • Author(s)
      Kinoshita M, Miyata T, Kawamoto A, Kato T, Namba K, Minamino T
    • Organizer
      第56回日本生物物理学会年会
    • Related Report
      2018 Research-status Report
  • [Presentation] Rotational symmetry structure of the bacterial flagellar motor for torque transmission revealed by electron cryomicroscopy (クライオ電子顕微鏡で解き明かす細菌べん毛モーターのトルク伝達に重要な回転対称構造)2018

    • Author(s)
      Kawamoto A, Miyata T, Kinoshita M, Minamino T, Imada K, Kato T, Namba K
    • Organizer
      第56回日本生物物理学会年会
    • Related Report
      2018 Research-status Report
  • [Presentation] Structure of the growing end of the bacterial flagellar filament (細菌べん毛繊維の成長端の構造)2018

    • Author(s)
      Kida M, Makino F, Kinoshita M, Miyata T, Kato T, Minamino T, Namba K, Imada K
    • Organizer
      第56回日本生物物理学会年会
    • Related Report
      2018 Research-status Report
  • [Remarks]

    • URL

      http://www.fbs.osaka-u.ac.jp/jpn/general/lab/02/

    • Related Report
      2019 Annual Research Report
  • [Remarks] 大阪大学大学院生命機能研究科日本電子YOKOGUSHI協働研究所 研究成果

    • URL

      http://www.fbs.osaka-u.ac.jp/jpn/general/lab/02/result/

    • Related Report
      2018 Research-status Report

URL: 

Published: 2018-04-23   Modified: 2021-02-19  

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