Understanding dynamics of the actin filament using cryoEM and structural determination from a small number of images

• Project/Area Number: 18H02410
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 43040:Biophysics-related
• Research Institution: Nagoya University
• Principal Investigator: Narita Akihiro 名古屋大学, 理学研究科, 准教授 (30360613)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥17,680,000 (Direct Cost: ¥13,600,000、Indirect Cost: ¥4,080,000); Fiscal Year 2020: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2019: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2018: ¥9,100,000 (Direct Cost: ¥7,000,000、Indirect Cost: ¥2,100,000)
• Keywords: 電子顕微鏡 / アクチン線維 / 細胞骨格 / アクチン動態 / 線維端 / アクチン / 走査透過型電子顕微鏡 / 単粒子解析 / 少数構造生物学 / アクチン線維ホモログ / クライオ電子顕微鏡

Outline of Final Research Achievements

High-resolution structural analysis of proteins that bind to actin molecules 1: 1 and actin filaments themselves has progressed, but analysis of actin fiber-binding proteins that have symmetry different from that of actin filaments remained to be done. In this study, we succeeded in one such target, the actin filament end, at 8 angstroms, three times higher than conventional one. In addition, by combining negatively stained images and scanning transmission electron microscopy, it became possible to analyze the properties of filaments from a small number of images, and three papers were published using this.

Academic Significance and Societal Importance of the Research Achievements

近年、クライオ電子顕微鏡による単粒子構造解析はめざましい発展を遂げたが、線維端やローカルな構造変化など、像の数を集めるのが難しい対象に対しては構造解析が進んでいない。この研究は、そのような難しい対象を構造解析のターゲットに取り込む研究であり、アクチン線維端について成功を収めた。また、少数の像から構造情報を取り出す技術を発展させることに成功、いくつかの論文を発表した。従来の高分解能構造解析とは異なる方向から、構造生物学の領域を広げることに貢献できたと考えている。

Research Products

• [Int'l Joint Research] Institut Curie(フランス)
• [Int'l Joint Research] A*STAR/National University of Singapore(シンガポール)
• [Journal Article] D-Loop Mutation G42A/G46A Decreases Actin Dynamics (2020)
  • Author(s): Matsuzaki Mizuki、Fujiwara Ikuko、Kashima Sae、Matsumoto Tomoharu、Oda Toshiro、Hayashi Masahito、Maeda Kayo、Takiguchi Kingo、Ma?da Yuichiro、Narita Akihiro
  • Journal Title: Biomolecules Volume: 10 Issue: 5 Pages: 736-736
  • DOI: 10.3390/biom10050736
  • Peer Reviewed / Open Access
• [Journal Article] The structure of a 15-stranded actin-like filament from Clostridium botulinum (2019)
  • Author(s): Koh Fujiet、Narita Akihiro、Lee Lin Jie、Tanaka Kotaro、Tan Yong Zi、Dandey Venkata P.、Popp David、Robinson Robert C.
  • Journal Title: Nature Communications Volume: 10 Issue: 1 Pages: 2856-2856
  • DOI: 10.1038/s41467-019-10779-9
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Helical rotation of the diaphanous-related formin mDia1 generates actin filaments resistant to cofilin. (2018)
  • Author(s): Mizuno, H., Tanaka, K., Yamashiro, S., Narita, A. & Watanabe, N.
  • Journal Title: Proc Natl Acad Sci U S A Volume: 115
  • NAID: 120006471210
  • Peer Reviewed
• [Presentation] アクチン線維構造と運動マシナリー (2020)
  • Author(s): 成田哲博
  • Organizer: 第43回分子生物学会年会
  • Invited
• [Presentation] 細胞運動のしくみ解明に挑戦する構造生物学 ～アクチン線維の精密解析～ (2020)
  • Author(s): 成田哲博
  • Organizer: 未来に挑戦する構造生物科学
  • Invited
• [Presentation] アクチン線維および関連タンパク質のクライオ電子顕微鏡法による高分解能構造解析 (2019)
  • Author(s): 成田哲博
  • Organizer: 岐阜構造生物学・医学・論理的創薬シンポジウム
  • Invited
• [Presentation] アクチン線維および関連タンパク質の高分解能構造解析 (2019)
  • Author(s): 成田哲博
  • Organizer: 構造生物学研究会 大阪大学
  • Invited
• [Presentation] High resolution structural analysis of the actin filaments and actin related proteins (2019)
  • Author(s): Akihiro Narita
  • Organizer: ”Frontiers in Cellular, Viral and Molecular Microscopy with Cryo-specimen Preparation Techniques
  • Int'l Joint Research / Invited
• [Presentation] Development of cryo-electron microscope for simultaneous STEM, SEMimaging (2018)
  • Author(s): Azuma J., Nagakubo Y., Tamba Y., Sato H., SunaoshiT., Tamochi R., Ose Y., Uuskura J., Narita A., Matsumoto T., Usukura E., Osumi M.:
  • Organizer: 第74回日本顕微鏡学会学術講演会
• [Presentation] アクチン線維構造解析と新しい電子顕微鏡法 (2018)
  • Author(s): 成田哲博
  • Organizer: 第79回応用物理学会秋季学術講演会
  • Invited