Elucidation of acute actin polymerization signaling using single molecule imaging of filament end factors

• Project/Area Number: 19390075
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: General medical chemistry
• Research Institution: Kyoto University
• Principal Investigator: WATANABE Naoki Kyoto University, 医学研究科, 准教授 (80303816)
• Project Period (FY): 2007 – 2008
• Project Status: Completed (Fiscal Year 2008)
• Budget Amount: ¥19,110,000 (Direct Cost: ¥14,700,000、Indirect Cost: ¥4,410,000); Fiscal Year 2008: ¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000); Fiscal Year 2007: ¥12,610,000 (Direct Cost: ¥9,700,000、Indirect Cost: ¥2,910,000)
• Keywords: 細胞 / 組織 / 一分子計測 / 生体分子 / ナノバイオ / 細胞・組織 / コフィリン / アクチン重合 / 細胞シグナル / フォルミンファミリー / アクチン線維切断

Research Abstract

多くの生命機能は細胞の形態変化と密接に連関し、動的なアクチン重合が多く関わっている。まず、アクチン脱重合因子AIP1 の単分子可視化により、細胞伸展部で毎秒280アクチン分子長の線維に1回線維が切断されることを見出した. これは主要なアクチン重合端形成機構と考えられたArp2/3複合体による重合核形成の15倍の高頻度である. また, フォルミン蛋白質mDia1がアクチン単量体増加にトリガーされ, プロセッシブに線維伸長させるアクチン線維修復機構を見出した. 線維崩壊から迅速にアクチン再重合させるフィードバック機構が明らかとなった。

Research Products

• [Journal Article] Cdc42 and Rac family GTPases regulate mode and speed but not direction of primary fibroblast migration during PDGF dependent chemotaxis (2009)
  • Author(s): Monypenny, J., Zicha, D., Higashida, C., Oceguera-Yanez, F., Narumiya, S. and Watanabe, N.
  • Journal Title: Mol.Cell. Biol. 29 Pages: 2730-2747
• [Journal Article] An order of magnitude faster AIP1 associated actin disruption than nucleation by the Arp2/3 complex in lamellipodia (2009)
  • Author(s): Tsuji, T., Miyoshi, T., Higashida, C.,Narumiya, S. Watanabe, N.
  • Journal Title: PLoS ONE 4
• [Journal Article] Imatinib mesylate (STI571)-induced cell edge translocation of kinase active and kinase defective Abelson kinase : requirements of myristoylation and src homology 3 domain (2009)
  • Author(s): Fujita, A., Shishido, T., Yuan, Y., Inamoto, E., Narumiya, S. and Watanabe, N.
  • Journal Title: Mol. Pharmacol 75 Pages: 75-84
• [Journal Article] Cdc42 and Rac family GTPases regulate mode and speed but not direction of primary fibroblast migration during PDGF-dependent chemotaxis. (2009)
  • Author(s): Monypenny, J., Zicha, D., Higashida, C, Oceguera-Yanez, F., Narumiya, S., Watanabe, N
  • Journal Title: Molecular and Cellular Biology
  • Peer Reviewed
• [Journal Article] An order of magnitude faster AIPl-associated actin disruption than nucleation by the Arp2/3 complex in lamellipodia. (2009)
  • Author(s): Tsuji, T., Miyoshi, T., Higashida, C, Narumiya, S., Watanabe, N.
  • Journal Title: PLoS ONE 4
  • Peer Reviewed
• [Journal Article] Imatinib mesylate (STI571)-induced cell edge translocation of kinase-active and kinase-defective Abelson kinase : requirements of myristoylation and srchomology 3 domain. (2009)
  • Author(s): Fujita, A., Shishido, T., Yuan, Y., Inamoto, E., Narumiya, S., Watanabe. N.
  • Journal Title: Molecular Pharmacology 75 Pages: 75-84
  • Peer Reviewed
• [Journal Article] G-actin regulates rapid induction of actin nucleation by mDia1 to restore cellular actin polymers (2008)
  • Author(s): Higashida, C., Suetsugu, S., Tsuji, T., Monypenny, J., Narumiya, S. Watanabe, N.
  • Journal Title: J. Cell Sci. 121 Pages: 3403-3412
• [Journal Article] Shootin1 interacts with actin retrograde flow and L1-CAM to promote axon utgrowth (2008)
  • Author(s): Shimada T, Toriyama M, Uemura1 K, Kamiguchi H, Sugiura T, Watanabe N, *Inagaki N.
  • Journal Title: J. Cell Biol. 181 Pages: 817-829
• [Journal Article] mDia2 induces the actin scaffold for the contractile ring and stabilizes its position during cytokinesis in NIH 3T3 cells (2008)
  • Author(s): Watanabe S, Ando Y, Yasuda S, Hosoya H, Watanabe N, Ishizaki T, *Narumiya S.
  • Journal Title: Mol.Biol. Cell 19 Pages: 2328-2338
• [Journal Article] G-actin regulates rapid induction of actin nucleation by mDial to restore cellular actin polymers. (2008)
  • Author(s): Higashida, C., Suetsugu, S., Tsuji, T., Monypenny, J., Narumiya, S., Watanabe, N.
  • Journal Title: Journal of Cell Science 121 Pages: 3403-3412
  • Peer Reviewed
• [Journal Article] Shootinl interacts with actin retrograde flow and LI-CAM to promote axon outgrowth. (2008)
  • Author(s): Shimada, T., Toriyama, M., Uemura, K., Kamiguchi, H., Sugiura, T., Watanabe. N., Inagaki, N.
  • Journal Title: Journal of Cell Biology 181 Pages: 817-829
  • Peer Reviewed
• [Journal Article] mDia2 induces the actin scaffold for the contractile ring and stabilizes its position during cytokinesis in NIH 3T3 cells. (2008)
  • Author(s): Watanabe, S., Ando, Y., Yasuda, S., Hosoya, H., Watanabe. N., Ishizaki, T., Narumiya, s.
  • Journal Title: Molecular Biology of the Cell 19 Pages: 2328-2338
  • Peer Reviewed
• [Journal Article] mDia2 Induces the Actin Scaffold for the Contractile Ring and Stabilizes its Position during Cytokinesis in NIH 3T3 Cells. (2008)
  • Author(s): Watanabe S., Ando Y., Yasuda S., Hosoya, H., Watanabe N., Ishizaki T. and Narumiya S.
  • Journal Title: Molecular Biology of the Cell (印刷中)
  • Peer Reviewed
• [Journal Article] アクチン重合システムを解く分子イメージング:みえない脱重合反応をとらえる (2007)
  • Author(s): 渡邊 直樹
  • Journal Title: 分子細胞治療 第6巻 Pages: 77-82
• [Journal Article] Shootinl interacts with actin retrograde flow and L1-CAM to promote axono utgrowth.
  • Author(s): Shimada T., Toriyama M., Uemura K., Kamiguchi H., Sugiura T, , Watanabe N. and Inagaki N.
  • Journal Title: Journal of Cell Biology (印刷中)
  • Peer Reviewed
• [Presentation] STI571 induced cell edge translocation of abl kinase (2009)
  • Author(s): Yuan Y., Fujita A., Shishido T., Narumiya S., Watanabe N
  • Organizer: 第82回日本薬理学会年会
  • Place of Presentation: 横浜(speaker)
  • Year and Date: 2009-03-18
• [Presentation] STI571に誘導されるablの細胞辺縁トランスロケーション (2009)
  • Author(s): Yuan Y., Fujita A., Shishido T., Narumiya S., Watanabe N. (speaker)
  • Organizer: 第82回日本薬理学会年会
  • Place of Presentation: 横浜
  • Year and Date: 2009-03-18
• [Presentation] A role of homeostasis between G- and F-actin in filament nucleation and actin polymer restoration by mDial. (2008)
  • Author(s): Higashida C., Watanabe N.
  • Organizer: 第48回アメリカ細胞生物学会
  • Place of Presentation: 米国サンフランシスコ
  • Year and Date: 2008-12-15
• [Presentation] Rapid actin polymer restoration mechanism by mDia1 and othe actin nucleators regulated by concentration fluctuation of G-actin (2008)
  • Author(s): Higashida C., Suetsugu S., Narumiya S., Watanabe N.
  • Organizer: 第31回日本分子生物学会年会・第81回日本生化学会大会合同大会シンポジウム(企画・主催)
  • Place of Presentation: 神戸(speaker)
  • Year and Date: 2008-12-11
• [Presentation] Rapid actin polymer restoration mechanism by mDial and other actin nucleators regulated by concentration fluctuation of G-actin. (2008)
  • Author(s): Higashida C., Suetsugu S., Narumiya S., Watanabe N. (speaker)
  • Organizer: 第31回日本分子生物学会年会・第81回日本生化学会大会合同大会(備考 : シンポジウム主催)
  • Place of Presentation: 神戸
  • Year and Date: 2008-12-11
• [Presentation] A paradoxical effect of actin monomer inhibitors to increase drug free G actin in cells : an unexpected role of G actin revealed by pharmacokinetic simulation (2008)
  • Author(s): Watanabe N., Higashida C.
  • Organizer: The Uehara Memorial Foundation Symposium 2008.; Systems Biology : the Challenge of Complexity
  • Place of Presentation: 東京
  • Year and Date: 2008-07-01
• [Presentation] A paradoxical effect of actin monomer inhibitors to increase drug-free G-actin in cells : an unexpected role of G-actiti revealed by pharmacokinetic simulation. (2008)
  • Author(s): Watanabe, N., Higashida, C.
  • Organizer: THE UEHARA MEMORIAL FOUNDATIONSYMPOSIUM-2008 Systems Biology : The Challenge of Complexity
  • Place of Presentation: 東京
  • Year and Date: 2008-07-01
• [Presentation] Challenging frequent filament severing annealing hypothesis by single molecule imaging of AIP1, a cofilin dependent actin barbed end interacting protein (2007)
  • Author(s): Watanabe N., Higashida C., Tsuji T.
  • Organizer: 第30回日本分子生物学会年会・第80回日本生化学会大8会合同大会ワークショップ(企画・主催)
  • Place of Presentation: 横浜
  • Year and Date: 2007-12-11
• [Presentation] Challenging frequent filament severing-annealing hypothesis by single-molecule imaging of AIP1, a cofilin-dependent actin barbed end interacting protein. (2007)
  • Author(s): Naoki Watanabe
  • Organizer: 第30回日本分子生物学会年会・第80回日本生化学会大会 合同大会
  • Place of Presentation: 横浜
  • Year and Date: 2007-12-11
• [Presentation] 細胞内G&Fアクチンのホメオスターシス (2007)
  • Author(s): 渡邊 直樹
  • Organizer: 「アクチン・ミオシン:分子と細胞を如何にして結び付けるか」の会(特定領域研究「生体ナノシステムの制御」主催)
  • Place of Presentation: 東京(早稲田大学)
  • Year and Date: 2007-11-10
• [Presentation] Molecular kinetics of AIP1, a cofilin-dependent actin barbed end interacting protein, in lamellipodia: Evidence for frequent filament severing. (2007)
  • Author(s): Naoki Watanabe
  • Organizer: SEMM Workshop on Cell Migration: from Molecules to Organisms and Diseases.
  • Place of Presentation: イタリア・ミラノ
  • Year and Date: 2007-05-13
• [Book] Systems Biology The Challenge of Complexity. Chapter : "A possible role of homeostasis between monomeric and filamentous actin in filament nucleation reveald by pharmacokinetic modeling. " (2009)
  • Author(s): Watanabe N., Higashida C.
  • Total Pages: 7
  • Publisher: Springer
• [Book] A possible role of homeostasis between monomeric and filamentous actin in filament nucleation reveald by pharmacokinetic modeling (2008)
  • Author(s): Watanabe N. and Higashida C.
  • Publisher: Systems Biology The Challenge of Complexity. Springer
• [Book] 非侵襲・可視化技術ハンドブックーナノ・バイオ・医療から情報システムまで-(共著)第8章第10節 アクチン細胞骨格のダイナミクスの可視化 (2007)
  • Author(s): 渡邊 直樹(小川誠二, 上野照剛編)
  • Total Pages: 8
  • Publisher: エヌ・ティー・エス出版
• [Remarks]
  • URL: http://www5.mfour.med.kyoto-u.ac.jp/n-watanabe.html
• [Remarks]
  • URL: http://www5.mfour.med.kyoto-u.ac.jp/n-watanabe.html