Single-molecule imaging analysis of cell structural reorganization processes downstream of G protein signaling

2009 Fiscal Year Final Research Report

• Project/Area Number: 17079004
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Kyoto University
• Principal Investigator: WATANABE Naoki Kyoto University, 医学研究科, 准教授 (80303816)
• Project Period (FY): 2005 – 2009
• Keywords: 細胞・組織 / 一分子計測 / G蛋白質 / フォルミンファミリー / Rhoファミリー

Research Abstract

Using fluorescence single-molecule imaging which directly visualizes remodeling processes of the actin cytoskeleton, this research project has revealed a novel actin polymer regeneration mechanism by which an increase in the cellular monomeric actin concentration provokes rapid actin nucleation by Formin homology proteins. This project has also quantitatively proved the frequent actin filament disruption in the cell. In addition, through various types of cell imaging analysis, the roles of Rho family GTPases in cell migration and inhibitor-induced conformational regulation of a protein kinase, Abl, have been elucidated.

Research Products

• [Journal Article] Inside view of cell locomotion through single-molecule: fast F-/G-actin cycle and G-actin regulation of polymer restoration. (2010)
  • Author(s): Watanabe, N.
  • Journal Title: Proc. Jpn. Acad. B. Phys. Biol. Sci. 86 Pages: 62-83
  • Peer Reviewed
• [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: Mol. Cell. Biol. 29 Pages: 2730-2747
  • Peer Reviewed
• [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 Pages: e4921
  • Peer Reviewed
• [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., Watanabe, N.
  • Journal Title: Mol. Pharmacol 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
  • Peer Reviewed
• [Journal Article] Shootin1 interacts with actin retrograde flow and L1-CAM to promote axon outgrowth. (2008)
  • Author(s): Shimada, T., Toriyama, M., Uemura, K., Kamiguchi, H., Sugiura, T., Watanabe, N., Inagaki, N.
  • Journal Title: J. Cell Biol. 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: Mol. Biol. Cell 19 Pages: 2328-2338
  • Peer Reviewed
• [Journal Article] Actin turnover-dependent fast dissociation of capping protein in the dendritic nucleation actin network: evidence of frequent filament severing. (2006)
  • Author(s): Miyoshi, T., Tsuji, T., Higashida, C., Hertzog, M., Fujita, A., Narumiya, S., Scita, G., Watanabe, N.
  • Journal Title: J. Cell Biol. 175 Pages: 947-955
  • Peer Reviewed
• [Presentation] 細胞内分子イメージングと低分子化合物研究の融合に向けて (2010)
  • Author(s): 渡邊直樹
  • Organizer: 第2回定量生物の会
  • Place of Presentation: 大阪
  • Year and Date: 2010-01-10
• [Presentation] A Role of G- and F-actin Homeostasis in Actin Polymer Restoration. (2009)
  • Author(s): Watanabe N., Higashida C., Mizuno H.
  • Organizer: Symposium on Watching Biomolecules in Action
  • Place of Presentation: 大阪
  • Year and Date: 2009-12-15
• [Presentation] Rapid actin polymer restoration against mechanical force: Roles of G- and F-actin homeostasis and mDia1. (2009)
  • Author(s): Watanabe N., Higashida C., Mizuno H.
  • Organizer: 第61回日本細胞生物学会大会
  • Place of Presentation: 名古屋
  • Year and Date: 2009-06-03
• [Presentation] STI571-induced cell edge translocation of abl kinase. (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] (speaker) Rapid actin polymer restoration mechanism by mDia1 and other 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
  • 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] 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] 細胞内G&Fアクチンのホメオスターシス「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」 (2007)
  • Author(s): 渡邊直樹
  • Organizer: 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] (○: speaker) Molecular kinetics of AIP1, a cofilin-dependent actin barbed end interacting protein, in lamellipodia: Evidence for frequent filament severing. (2007)
  • Author(s): Tsuji T., Higashida C., ○Watanabe N.
  • 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
• [Presentation] Lessons from single-molecule microscopy of cellular actin systems. Part1: frequent severing-annealing of filaments. (2007)
  • Author(s): Watanabe N., Higashida C. (joint speaker)
  • Organizer: Part 2: acute filament restoration by mDia1. 英国癌研究所(Cancer Research UK)
  • Place of Presentation: イギリスロンドン
  • Year and Date: 2007-05-10
• [Presentation] Acute actin polymerization mechanisms by mDia1 in living cells. (2006)
  • Author(s): 渡邊直樹
  • Organizer: 日本分子生物学会2006フォーラム
  • Place of Presentation: 名古屋
  • Year and Date: 2006-12-06
• [Presentation] 単分子イメージングによる細胞内アクチン重合機構の解析 (2005)
  • Author(s): 渡邊直樹
  • Organizer: 第14回日本バイオイメージング学会学術集会
  • Place of Presentation: 東京
  • Year and Date: 2005-10-27
• [Presentation] Evidence of constitutive actin filament severing in lamellipodia. (2005)
  • Author(s): Miyoshi T., Higashida C., ○Watanabe N. (○: speaker)
  • Organizer: 第58回日本細胞生物学会大会
  • Place of Presentation: 埼玉県大宮
  • Year and Date: 2005-06-17
• [Presentation] Molecular Kinetics of the Arp 2/3 Complex and Capping Proteins at the Leading Edge. (2005)
  • Author(s): Miyoshi T., Higashida C., ○Watanabe N. (○: speaker)
  • Organizer: Keystone Symposia, Cell Migration and Adhesion
  • Place of Presentation: 日米国ユタ州スノーバード
  • Year and Date: 2005-04-10
• [Book] Nat. Cell Biol. 11(Migration without a clutch.) (2009)
  • Author(s): Narumiya, S., Watanabe, N.
  • Total Pages: 1394-1396
  • Publisher: News and Views
• [Book] A possible role of homeostasis between monomeric and filamentous actin in filament nucleation revealed by pharmacokinetic modeling. Systems Biology The Challenge of Complexity (2009)
  • Author(s): Watanabe, N., Higashida, C.
  • Total Pages: 143-149
  • Publisher: Springer
• [Book] アクチン細胞骨格のダイナミクスの可視化非侵襲・可視化技術ハンドブック-ナノ・バイオ・医療から情報システムまで- (2007)
  • Author(s): 渡邊直樹
  • Total Pages: 843-850
  • Publisher: NTS出版
• [Book] 生理活性物質のケミカルバイオロジーを開拓した論文(Mayer, T.U. et. al. ) (2007)
  • Author(s): 木下専, 渡邊直樹
  • Total Pages: 971-974,1796-1799
• [Book] 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分子細胞治療 (2007)
  • Author(s): 渡邊直樹
  • Total Pages: 77-82
• [Book] 単分子イメージングが明らかにした生細胞内アクチン重合機構日本臨床 (2007)
  • Author(s): 辻貴宏, 渡邊直樹
  • Total Pages: 235-241
• [Book] 単分子スペックル顕微鏡によるアクチンダイナミックス観察実 験医学 (2006)
  • Author(s): 渡邊直樹
  • Total Pages: 186-193
• [Book] アクチン細胞骨格とアクチン調節タンパク質実験医学第24巻第13号増刊形と運動を司る細胞のダイナミクス (2006)
  • Author(s): 馬渕一誠, 渡邊直樹, 寺崎朝子, 高稲正勝, 斧正一郎
  • Total Pages: 64-75
• [Book] アクチンダイナミクス蛋白質核酸酵素 (2006)
  • Author(s): 渡邊直樹
  • Total Pages: 522-528
• [Book] アクチン重合が駆動する細胞運動の謎に挑む単分子イメージング生物物理 (2005)
  • Author(s): 渡邊直樹, 東田知陽, 三好拓志
  • Total Pages: 292-296
• [Book] 単分子スペックル顕微鏡によるアクチン生化学の一分子イメージング蛋白質核酸酵素 (2005)
  • Author(s): 東田知陽, 三好拓志, 渡邊直樹
  • Total Pages: 1436-1442
• [Book] mDia1とForminファミリー:アクチン伸長端をサーフィンするプロセッシブキャッパー生化学 (2005)
  • Author(s): 渡邊直樹, 東田知陽
  • Total Pages: 136-140
• [Book] 単分子スペックル顕微鏡がみせるアクチン重合の細胞内分子キネティクス日本薬理学雑誌 (2005)
  • Author(s): 渡邊直樹
  • Total Pages: 103-108
• [Book] Forminファミリー (2005)
  • Author(s): 渡邊直樹, 東田知陽
  • Total Pages: 150-152
• [Book] 細胞の中を分子でのぞく-Less is moreとSerendipity-学術月報 (2005)
  • Author(s): 渡邊直樹
  • Total Pages: 389-391
• [Remarks]
  • URL: http://www.lifesci.tohoku.ac.jp/outline/biomolecular/single-molecule.html