Translocation pathway of the bacterial flagellar typeIII secretion system analyzed by site-directed photo-crosslinking

• Project/Area Number: 18K07108
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 49050:Bacteriology-related
• Research Institution: Nagasaki University (2020); Nagoya University (2018-2019)
• Principal Investigator: TERASHIMA Hiroyuki 長崎大学, 熱帯医学研究所, 助教 (60791788)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2020: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000); Fiscal Year 2019: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2018: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: 細菌べん毛 / III型分泌装置 / 光架橋 / in vitro再構成 / タンパク質分泌 / 輸送装置 / タンパク質輸送 / 反転膜

Outline of Final Research Achievements

We studied the mechanism of protein export, construction and function of the bacterial flagellum. Using an in vitro transport assay with the inverted membrane vesicles, we were able to construct hook and filament in vitro. The length of hook was regulated to 55 nm by the molecular ruler protein FliK. These results revealed that the flagellum is a molecular machine that is autonomously constructed using only flagellar components.

Academic Significance and Societal Importance of the Research Achievements

細菌べん毛は、細胞外に長く伸びた線維状の超分子複合体である。その構築メカニズムを明らかにすることによって、タンパク質から構築される分子機械の作動原理と設計原理を知ることができる。本研究の成果によって、べん毛の構築原理の一端を解き明かすことができ、学術的意義のある研究成果を得ることができた。

Research Products

• [Journal Article] Site-directed crosslinking identifies the stator-rotor interaction surfaces in a hybrid bacterial flagellar motor (2021)
  • Author(s): Terashima Hiroyuki、Kojima Seiji、Homma Michio
  • Journal Title: Journal of Bacteriology Volume: 9 Issue: 9
  • DOI: 10.1128/jb.00016-21
  • Peer Reviewed
• [Journal Article] Assembly mechanism of a supramolecular MS-ring complex to initiate bacterial flagellar biogenesis in Vibrio species. (2020)
  • Author(s): Terashima H, Hirano K, Inoue Y, Tokano T, Kawamoto A, Kato T, Yamaguchi E, Namba K, Uchihashi T, Kojima S, Homma M.
  • Journal Title: Journal of Bacteriology Volume: - Issue: 16
  • DOI: 10.1128/jb.00236-20
  • Peer Reviewed
• [Journal Article] In Vitro Autonomous Construction of the Flagellar Axial Structure in Inverted Membrane Vesicles. (2020)
  • Author(s): Terashima H, Tatsumi C, Kawamoto A, Namba K, Minamino T, Imada K.
  • Journal Title: Biomolecules Volume: 10 Issue: 1 Pages: 126-126
  • DOI: 10.3390/biom10010126
  • Peer Reviewed / Open Access
• [Journal Article] Structure of Vibrio FliL, a New Stomatin-like Protein That Assists the Bacterial Flagellar Motor Function (2019)
  • Author(s): Norihiro Takekawa, Miyu Isumi, Hiroyuki Terashima, Shiwei Zhu, Yuuki Nishino, Mayuko Sakuma, Seiji Kojima, Michio Homma, Katsumi Imada
  • Journal Title: mBio Volume: 10(2) Issue: 2
  • DOI: 10.1128/mbio.00292-19
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] In Vitro Reconstitution of Functional Type III Protein Export and Insights into Flagellar Assembly (2018)
  • Author(s): Hiroyuki Terashima, Akihiro Kawamoto, Chinatsu Tatsumi, Keiichi Namba, Tohru Minamino, Katsumi Imada
  • Journal Title: mBio Volume: 9(3) Issue: 3
  • DOI: 10.1128/mbio.00988-18
  • Peer Reviewed / Open Access
• [Journal Article] Novel insight into an energy transduction mechanism of the bacterial flagellar type III protein export (2018)
  • Author(s): Hiroyuki Terashima, Katsumi Imada
  • Journal Title: Biophysics and Physicobiology Volume: 15 Issue: 0 Pages: 173-178
  • DOI: 10.2142/biophysico.15.0_173
  • NAID: 130007437793
  • ISSN: 2189-4779
  • Peer Reviewed / Open Access
• [Presentation] 細菌べん毛回転に必須な固定子‐回転子間の相互作用機序 (2021)
  • Author(s): 〇寺島浩行、小嶋誠司、本間道夫
  • Organizer: 第94回日本細菌学会総会
  • Invited
• [Presentation] In vitro分泌再構成系を用いたべん毛III型分泌装置の作動メカニズムの解明 (2020)
  • Author(s): 〇寺島 浩行、川本 晃大、巽 千夏、難波 啓一、南野 徹、本間 道夫、今田 勝巳
  • Organizer: 第34回日本バイオフィルム学会学術集会&第57回日本細菌学会中部支部総会
• [Presentation] 部位特異的架橋法による細菌べん毛固定子－回転子間相互作用の検出 (2020)
  • Author(s): 〇寺島浩行、小嶋誠司、本間道夫
  • Organizer: 日本生体エネルギー研究会 第46回討論会
• [Presentation] 網羅的光架橋法による細菌べん毛回転モーターの固定子ー回転子間相互作用の解析 (2020)
  • Author(s): 〇寺島浩行、小嶋誠司、本間道夫
  • Organizer: 第93回日本生化学会大会
• [Presentation] 細菌III型分泌装置に対する阻害剤を探索だい929するための新規網羅的アッセイ系 (2019)
  • Author(s): 〇寺島浩行、川本晃大、巽千夏、難波啓一、南野徹、本間道夫、今田勝巳
  • Organizer: 第92回日本生化学会大会
  • Invited
• [Presentation] 細菌べん毛輸送装置によるタンパク質輸送の反転膜小胞を用いたin vitro再構成 (2019)
  • Author(s): ○寺島浩行、川本晃大、巽千夏、難波啓一、南野徹、今田勝巳
  • Organizer: 第83回日本生化学会中部支部例会