バクテリア巨大渦状運動の極性形成メカニズム

• Project Area: Synergy of Fluctuation and Structure:Foundation of Universal Laws in Nonequilibrium Systems
• Project/Area Number: 16H00808
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Science and Engineering
• Research Institution: Gakushuin University
• Principal Investigator: 西坂 崇之 学習院大学, 理学部, 助教授 (40359112)
• Project Period (FY): 2016-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥10,140,000 (Direct Cost: ¥7,800,000、Indirect Cost: ¥2,340,000); Fiscal Year 2017: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2016: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000)
• Keywords: 滑走運動 / 線毛 / 集団運動 / バイオフィルム / フラボバクテリウム・ジョンソニエ / 渦状運動

Outline of Annual Research Achievements

本課題は申請段階において、フラボバクテリウム・ジョンソニエという一種類の桿菌を研究対象にして開始された。その後の２年間の研究期間において、シネコシスティス、ハロバクテリウム、サーマス・サーモフィラスといったさまざまな研究対象に派生し、その幾つかは既に原著論文として発表されている。
ミキソコッカスのようなバクテリアで見られる、複数の個体の固まりが方向性を持って動くsocial motility（S-motility）は、特徴的な接着装置であるIV型線毛（type IV pili）によって実現される。このpiliのダイナミクスには膜内外のさまざまなタンパク質が関わるものの、個々の役割や詳細な機能については、まだ解明が進んでいないというのが現状である。本課題では、独自の光学顕微鏡を駆使することによって、piliの活性がどのように制御され、そして方向性のある運動を引き起こすのかについて、徹底的な可視化を進めた。
最終年度では、特にサーマス・サーモフィラスをモデル生物とし、１個体の運動を定量的に解析した。走光性という特徴に着目し、変異株を利用しながら制御の仕組みについて調べ、未踏の領域である「トゥイチング運動の制御機構」に関する本質的な知見を得ることができた。また繊維状のタンパク質であるPilAのダイナミクスについては、先行研究においても、実験条件の難しさから可視化は実現されていない。蛍光蛋白や微小粒子などを通じて、PilAの伸長と短縮を画像化し、その動態を直接観察した。

Research Progress Status

29年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

29年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] Detailed analyses of stall force generation in Mycoplasma mobile gliding. (2018)
  • Author(s): Mizutani M, Tulum I, Kinosita Y, Nishizaka T and Miyata M
  • Journal Title: Biophysical Journal Volume: 114 Issue: 6 Pages: 1411-9
  • DOI: 10.1016/j.bpj.2018.01.029
  • Peer Reviewed / Open Access
• [Journal Article] Unforeseen swimming and gliding mode of an insect gut symbiont, Burkholderia sp. RPE64, with wrapping of the flagella around its cell body (2017)
  • Author(s): Kinosita Yoshiaki、Kikuchi Yoshitomo、Mikami Nagisa、Nakane Daisuke、Nishizaka Takayuki
  • Journal Title: The ISME Journal Volume: 12 Issue: 3 Pages: 838-848
  • DOI: 10.1038/s41396-017-0010-z
  • Peer Reviewed
• [Journal Article] Asymmetric distribution of type IV pili triggered by directional light in unicellular cyanobacteria (2017)
  • Author(s): Nakane Daisuke、Nishizaka Takayuki
  • Journal Title: Proceedings of the National Academy of Sciences Volume: 114 Issue: 25 Pages: 6593-6598
  • DOI: 10.1073/pnas.1702395114
  • Peer Reviewed / Open Access
• [Journal Article] Dissection of the angle of single fluorophore attached to the nucleotide in corkscrewing microtubules (2017)
  • Author(s): Fujimura, S. Ito, Y. Ikeguchi, M. Adachi, K. Yajima, J. Nishizaka, T.
  • Journal Title: Biochem Biophys Res Commun Volume: 485 Issue: 3 Pages: 614-620
  • DOI: 10.1016/j.bbrc.2017.01.165
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Direct observation of rotation and steps of the archaellum in the swimming halophilic archaeon Halobacterium salinarum (2016)
  • Author(s): Kinosita, Y. Uchida, N. Nakane, D. Nishizaka, T.
  • Journal Title: Nat Microbiol Volume: 1 Issue: 11 Pages: 16148-16148
  • DOI: 10.1038/nmicrobiol.2016.148
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] F1-ATPase conformational cycle from simultaneous single-molecule FRET and rotation measurements (2016)
  • Author(s): Sugawa, M. Okazaki, K. I. Kobayashi, M. Matsui, T. Hummer, G. Masaike, T. Nishizaka, T.
  • Journal Title: Proc Natl Acad Sci USA Volume: 0 Issue: 21 Pages: 0-0
  • DOI: 10.1073/pnas.1524720113
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Directed binding of gliding bacterium, Mycoplasma mobile, shown by detachment force and bond lifetime (2016)
  • Author(s): Tanaka A, Nakane D, Mizutani M, Nishizaka T, Miyata M
  • Journal Title: mBio Volume: 7 Issue: 3 Pages: 1-1
  • DOI: 10.1128/mbio.00455-16
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] アーキアラとIV型線毛のモーターの動態を可視化する (2017)
  • Author(s): 西坂崇之
  • Organizer: 日本物理学会 第72回年次大会
  • Place of Presentation: 大阪大学 豊中キャンパス
  • Year and Date: 2017-03-17
  • Invited
• [Presentation] Unitary steps of supermolecular motility machineries in gliding bacteria and swimming archaea (2017)
  • Author(s): Takayuki Nishizaka
  • Organizer: Bacterial Flagella, Injectisomes and Type III Secretion Systems
  • Place of Presentation: Okinawa Institute of Science and Technology Graduate University
  • Year and Date: 2017-03-01
  • Int'l Joint Research / Invited
• [Presentation] "Development of optical-microscope techniques for single-molecule biophysics and their application to microbiology." (2017)
  • Author(s): Nishizaka, T.
  • Organizer: 10th International Conference on Advanced Materials and Devices
  • Int'l Joint Research / Invited
• [Presentation] "Development of Imaging Techniques to Reveal Singl-Protein Dynamics and their Application to Microbiology." (2017)
  • Author(s): Nishizaka, T.
  • Organizer: International Symposium on Fluctuation and Structure out of Equilibrium 2017
  • Int'l Joint Research / Invited
• [Presentation] “Development of Techniques for Single-Molecule Detection of Protein Conformational Dynamics and Their Application to Microbiology.” (2017)
  • Author(s): Nishizaka, T.
  • Organizer: The 2nd Korea-Japan Joint Symposium on Single-Molecule Biophysics
  • Int'l Joint Research / Invited
• [Presentation] "Application of single-molecule techniques to supramolecular machinery in bacteria and archaea." (2017)
  • Author(s): Nishizaka, T.
  • Organizer: The 15th International Conference of “Na,K-ATPases and Related Transport ATPases”
  • Int'l Joint Research / Invited
• [Presentation] "Unitary steps of supermolecular motility machineries in gliding bacteria and swimming archaea." (2017)
  • Author(s): Nishizaka, T.
  • Organizer: Bacterial Flagella, Injectisomes and Type III Secretion Systems
  • Int'l Joint Research / Invited
• [Presentation] Structural and functional dynamics of nanoscale molecular motors (2016)
  • Author(s): T. Nishizaka Y. Kinosita T. A. Katoh D. Nakane
  • Organizer: 31st International Congress on High Speed Imaging and Photonics
  • Place of Presentation: Hotel Hankyu Expo Park
  • Year and Date: 2016-11-07
  • Int'l Joint Research / Invited
• [Presentation] Visualization of dynamics of motors in archaella and type IV pili system (2016)
  • Author(s): Takayuki Nishizaka
  • Organizer: Current and Future Perspectives in Active Matter
  • Place of Presentation: The University of Tokyo
  • Year and Date: 2016-10-28
  • Invited
• [Presentation] 光学顕微鏡技術の新展開：膜超分子モーターへの先進的な応用 シンポジウム「膜タンパク分子作動原理解明の新基軸」 (2016)
  • Author(s): 西坂崇之
  • Organizer: 第89回日本生化学会大会
  • Place of Presentation: 東北大学川内北キャンパス
  • Year and Date: 2016-09-25
  • Invited
• [Book] Handbook of Photonics for Biomedical Engineering (2017)
  • Author(s): Katoh, T. A. Fujimura, S. Nishizaka, T.
  • Publisher: Springer
  • ISBN: 9789400750531