Mechanisms of actin polymerization, ATP hydrolysis, and filament severing revealed by F-form crystal structures

• Project/Area Number: 16K14708
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Biophysics
• Research Institution: Nagoya University
• Principal Investigator: Takeda Shuichi 名古屋大学, 理学研究科, 研究員 (50509081)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000); Fiscal Year 2017: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2016: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
• Keywords: アクチン / X線結晶構造解析 / フラグミン / ゲルゾリンスーパーファミリー / ATP加水分解 / 繊維切断 / 構造生物学 / ゲルゾリン

Outline of Final Research Achievements

The crystal structure of four actin subunits in complex with fragmin, a gelsolin superfamily protein, was solved at 2.3 A resolution. In addition, the crystal structures of single actin molecule in complex with a fragmin N-terminal domain, adopting the filamentous conformation, were determined at 1.2 A resolution. These F-form actin structures have made it possible to discuss the mechanism of actin polymerization and ATP hydrolysis at atomic resolution.

Academic Significance and Societal Importance of the Research Achievements

アクチンは不定長に重合するタンパク質であるため、これまで繊維状態での結晶構造は報告されていなかった。今回アクチン切断因子であるフラグミンとの共結晶化という方法によって、世界初のアクチン繊維構造の解明に成功した。本研究の成果は真核細胞中に最も大量に存在するタンパク質であるアクチンの機能を理解する上で極めて重要である。

Research Products

• [Journal Article] Polymerization and depolymerization of actin with nucleotide states at filament ends (2018)
  • Author(s): Fujiwara Ikuko、Takeda Shuichi、Oda Toshiro、Honda Hajime、Narita Akihiro、Ma?da Yuichiro
  • Journal Title: Biophysical Reviews Volume: 10 Issue: 6 Pages: 1513-1519
  • DOI: 10.1007/s12551-018-0483-7
  • Peer Reviewed
• [Journal Article] Structural basis for cofilin binding and actin filament disassembly (2018)
  • Author(s): Tanaka, K., Takeda, S., Mitsuoka, K., Oda, T., Kimura-Sakiyama, C., Maeda, Y., Narita, A.
  • Journal Title: Nat Commun Volume: 印刷中
  • Peer Reviewed / Open Access
• [Journal Article] Comprehensive analysis of motions in molecular dynamics trajectories of the actin capping protein and its inhibitor complexes (2016)
  • Author(s): Koike, R., Takeda, S., Maeda, Y., Ota, M.
  • Journal Title: Proteins Volume: 未定 Issue: 7 Pages: 948-956
  • DOI: 10.1002/prot.25043
  • Peer Reviewed
• [Presentation] BL2S1を利用したX線結晶構造解析から明らかとなったアクチン重合・ATP加水分解機構 (2019)
  • Author(s): 武田修一、成田哲博、小田俊郎、小池亮太郎、森次圭、太田元規、兼松佑典、藤原郁子、田中康太郎、永江峰幸、渡邉信久、前田雄一郎
  • Organizer: 第8回名古屋大学シンクロトロン光研究センターシンポジウム
  • Invited
• [Presentation] Atomic resolution structures of F‐form actin: mutual switching between G/F transition and ATPase (2018)
  • Author(s): Shuichi Takeda, Akihiro Narita, Toshiro Oda, Nobuhisa Watanabe, Takayuki Nagae, Motonori Ota, Ryotaro Koike, Kei Moritsugu, Yusuke Kanematsu, Ikuko Fujiwara, Kotaro Tanaka, Yuichiro Maeda
  • Organizer: The 47th European muscle conference
  • Int'l Joint Research / Invited
• [Presentation] The structure of F-form actin at atomic resolutions: mutual switching between polymorphic transition and ATPase (2018)
  • Author(s): Shuichi Takeda, Akihiro Narita, Toshiro Oda, Nobuhisa Watanabe, Y, Takayuki Nagae, Motonori Ota, Ryotaro Koike, Kei Moritsugu, Yusuke Kanematsu, Ikuko Fujiwara, Kotaro Tanaka, Yuichiro Maeda
  • Organizer: Myofilament conference 2018
  • Int'l Joint Research
• [Presentation] F-form crystal structures: Mechanisms of actin assembly and F-actin ATPase (2018)
  • Author(s): Shuichi Takeda, Akihiro Narita, Toshiro Oda, Kotaro Tanaka, Ryotaro Koike, Motonori Ota, Ikuko Fujiwara, Nobuhisa Watanabe, Yuichiro Maeda
  • Organizer: The 62th Annual Meeting of Biophysical Society
  • Int'l Joint Research
• [Presentation] F型アクチンの結晶構造とアクチン集合メカニズム (2017)
  • Author(s): 武田修一、前田雄一郎
  • Organizer: 生体運動研究合同班会議
  • Place of Presentation: 神戸国際会議場, 兵庫県神戸市
  • Year and Date: 2017-01-06
• [Presentation] ATPase mechanism and dynamic assembly of actin revealed by the F-form crystal structures (2017)
  • Author(s): 武田 修一、成田 哲博、小田俊郎、田中 康太郎、小池 亮太郎、太田 元規、藤原 郁子、渡邉 信久、前田 雄一郎
  • Organizer: 第55回日本生物物理学会年会
  • Invited
• [Presentation] Actin-cofilin complex structure at 3.8 A resolution revealed by cryo-EM (2017)
  • Author(s): 田中 康太郎、武田 修一、光岡 薫、小田 俊郎、前田 雄一郎、成田 哲博
  • Organizer: 第55回日本生物物理学会年会
• [Presentation] 高分解能X線結晶構造から明らかとなったアクチン重合・ATP加水分解機構 (2017)
  • Author(s): 武田 修一、成田 哲博、小田 俊郎、田中 康太郎、小池 亮太郎、太田 元規、藤原 郁子、渡邉 信久、前田 雄一郎
  • Organizer: 平成29年度日本結晶学会年会
• [Presentation] F型アクチンの結晶構造 (2017)
  • Author(s): 武田修一、成田哲博、小田俊朗、田中康太郎、渡邉信久、前田雄一郎
  • Organizer: 名古屋大学シンクロトロン光研究センターシンポジウム
  • Place of Presentation: 名古屋大学, 愛知県名古屋市
• [Presentation] Crystal Structure of the “F-form” Actin in Complex with Fragmin (2016)
  • Author(s): Shuichi Takeda, Akihiro Narita, Kotaro Tanaka, Toshiro Oda, Nobuhisa Watanabe, Yuichiro Maeda
  • Organizer: Now in actin study: Motor protein research reaching a new stage
  • Place of Presentation: 名古屋大学, 愛知県名古屋市
  • Year and Date: 2016-12-12
  • Int'l Joint Research / Invited
• [Presentation] Crystal Structure of Actin 4-mer in Complex with 2 Fragmin Molecules and the Mechanism of Actin Filament Assembly (2016)
  • Author(s): Takeda, S., Narita, A., Oda, T., Tanaka, K., Maeda, Y.
  • Organizer: Gordon Research Conference, Muscle & Molecular Motors
  • Place of Presentation: Mount Snow, West Dover, Vermont, USA
  • Year and Date: 2016-07-17
  • Int'l Joint Research / Invited