Molecular mechanisms of mitotic spindle organization and positioning

2011 Fiscal Year Final Research Report

• Project/Area Number: 21370086
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Cell biology
• Research Institution: Tohoku University
• Principal Investigator: MIZUNO Kensaku 東北大学, 大学院・生命科学研究科, 教授 (70128396)
• Project Period (FY): 2009 – 2011
• Keywords: コフィリン / 紡錘体 / 細胞分裂 / 細胞極性 / アクチン骨格

Research Abstract

We investigated the mechanisms of mitotic spindle organization and positioning and obtained the following results. 1)Plk1 binds to and phosphorylates cofilin-phosphatase Slingshot during mitosis. 2)NDR kinase and its upsteam regulators, MST and Furry, are crucial for bipolar spindle organization and chromosome alignment in metaphase. 3)Furry is involved in the maintenance of spindle bipolarity by promoting Aurora A-mediated Plk1 activation on spindle poles.

Research Products

• [Journal Article] Visualization of cofilin-actin and Ras-Raf interactions by bimolecular fluorescence complementation assays using a new pair of split Venus fragments (2012)
  • Author(s): Ohashi, K., Kiuchi, T., Shoji, K., Sampei, K., and Mizuno, K.
  • Journal Title: BioTechniques Volume: 52 Pages: 45-50
  • Peer Reviewed
• [Journal Article] Live-cell imaging of G-actin dynamics using sequential FDAP (2011)
  • Author(s): Kiuchi, T., Nagai, T., Ohashi, K., Watanabe, N., and Mizuno, K.
  • Journal Title: BioArchitecture Volume: 1 Pages: 240-244
  • Peer Reviewed
• [Journal Article] Cofilin-mediated F-actin severing is regulated by the Rap GTPase and controls the cytoskeletal dynamics that drive lymphocyte spreading and BCR microcluster formation (2011)
  • Author(s): Freeman, S. A., Lei, V., Dang-Lawson, M., Mizuno, K., Roskelley, C. D., and Gold, M. R.
  • Journal Title: J. Immunol Volume: 187 Pages: 5887-5900
  • Peer Reviewed
• [Journal Article] LIM-kinase has a dual role in regulating lamellipodium extension by decelerating the rate of actin retrograde flow and the rate of actin polymerization (2011)
  • Author(s): Ohashi, K., Fujiwara, S., Watanabe, T., Kondo, H., Kiuchi, T., Sato, M., and Mizuno, K.
  • Journal Title: J. Biol. Chem Volume: 286 Pages: 36340-36351
  • Peer Reviewed
• [Journal Article] Protein kinase D regulates cofilin activity through p21-activated kinase 4 (2011)
  • Author(s): Spratley, S. J., Bastea, L. I., Doppler, H., Mizuno, K., and Storz, P.
  • Journal Title: J. Biol. Chem Volume: 286 Pages: 34254-34261
  • Peer Reviewed
• [Journal Article] Measurements of spatiotemporal changes in G-actin concentration reveal its effect on stimulus-induced actin assembly and lamellipodium extension (2011)
  • Author(s): Kiuchi, T., Nagai, T., Ohashi, K., and Mizuno, K.
  • Journal Title: J. Cell Biol Volume: 193 Pages: 365-380
  • Peer Reviewed
• [Journal Article] CAMP (C13orf8, ZNF828)is a novel regulator of kinetochore-microtubule attachment (2011)
  • Author(s): Itoh, G., Kanno, S., Uchida, K. S. K., Chiba, S., Sugino, S., Watanabe, K., Mizuno, K., Yasui, A., Hirota, T., and Tanaka, K.
  • Journal Title: EMBO J Volume: 30 Pages: 130-144
  • Peer Reviewed
• [Journal Article] Global phosphorylation analysis of beta-arrestin-mediated signaling downstream of a seven transmembrane receptor (7TMR) (2010)
  • Author(s): Xiao, K., Sun, J., Kim, J., Rajagopal, S., Zhai, B., Villen, J., Haas, W., Kovacs, J. J., Shukla, A. K., Hara, M., Hernandez, M., Lachmann, A., Zhao, S., Lin, Y., Cheng, Y., Mizuno, K., Ma' ayan, A., Gygi, S. P., and Lefkowitz, R. J.
  • Journal Title: Proc Natl. Acad. Sci. U. S. A Volume: 107 Pages: 15299-15304
  • Peer Reviewed
• [Journal Article] Involvement of p114-RhoGEF and Lfc in Wnt-3a-and Dishevelled-induced RhoA activation and neurite retraction in N1E-115 mouse neuroblastoma cells (2010)
  • Author(s): Tsuji, T., Ohta, Y., Kanno, Y., Hirose, K., Ohashi, K., and Mizuno, K.
  • Journal Title: Mol. Biol. Cell Volume: 21 Pages: 3590-3600
  • Peer Reviewed
• [Journal Article] Actin depolymerizing factor cofilin-1 is necessary in maintaining mature podocyte architecture (2010)
  • Author(s): Garg, P., Verma, R., Cook, L., Soofi, A., Venkatareddy, M., George, B., Mizuno, K., Gurniak, c., Witke, W., and Holzman, L. B.
  • Journal Title: J. Biol. Chem Volume: 285 Pages: 22676-22688
  • Peer Reviewed
• [Journal Article] LIM-kinase is critical for the mesenchymal-to-amoeboid cell morphological transition in 3D matrices (2010)
  • Author(s): Mishima, T., Naotsuka, M., Horita, Y., Sato, M., Ohashi, K., and Mizuno, K.
  • Journal Title: Biochem. Biophys. Res. Commun Volume: 392 Pages: 577-581
  • Peer Reviewed
• [Journal Article] Requirements for recruitment of a G protein-coupled receptor to clathrin-coated pits in budding yeast (2009)
  • Author(s): Toshima, J. Y., Nakanishi, J., Mizuno, K., Toshima, J., and Drubin, D. G.
  • Journal Title: Mol. Biol. Cell Volume: 20 Pages: 5039-5050
  • Peer Reviewed
• [Journal Article] Ca2+/calmodulin-dependent protein kinase IV-mediated LIM-kinase activation is critical for calcium signal-induced neurite outgrowth (2009)
  • Author(s): Takemura, M., Mishima, T., Wang, Y., Kasahara, J., Fukunaga, K., Ohashi, K., and Mizuno, K.
  • Journal Title: J. Biol. Chem Volume: 284 Pages: 28554-28562
  • Peer Reviewed
• [Journal Article] MST2-and Furry-mediated activation of NDR1 is critical for precise alignment of mitotic chromosomes (2009)
  • Author(s): Chiba, S., Ikeda, M., Katsunuma, K., Ohashi, K., and Mizuno, K.
  • Journal Title: Curr. Biol Volume: 19 Pages: 675-681
  • Peer Reviewed
• [Journal Article] Protein kinase D1 regulates cofilin-mediated F-actin reorganization and cell motility through Slingshot (2009)
  • Author(s): Eiseler, T., Doppler, H., Yan, I. K., Kitatani, K., Mizuno, K., and Storz, P.
  • Journal Title: Nat. Cell Biol Volume: 11 Pages: 545-556
  • Peer Reviewed
• [Presentation] ラメリポディア伸長時におけるアクチンモノマー濃度の役割 (2012)
  • Author(s): 木内泰、渡邊直樹、水野健作
  • Organizer: 2012年生体運動研究合同班会議
  • Place of Presentation: 筑波大学
  • Year and Date: 20120106-08
• [Presentation] Furry is involved in bipolar spindle formation through Plk1 activation (2011)
  • Author(s): Ikeda, M., Chiba, S., Ohashi, K., and Mizuno K.
  • Organizer: 第34回日本分子生物学会
  • Place of Presentation: 横浜
  • Year and Date: 20111213-1216
• [Presentation] Live-cell measurements of spatiotemporal changes in G-actin concentration reveal its key role in stimulus-induced actin assembly and cell extension (2011)
  • Author(s): Mizuno, K.
  • Organizer: The 26th European Cytoskeletal Forum (ECF)Meeting on"Actin-based motility : from molecules to model organisms"
  • Place of Presentation: Stresa, Lake Maggiore, Italy
  • Year and Date: 20111029-1102
• [Presentation] Regulation mechanisms of actin cytoskeleton and membrane transport for the formation of cellular protrusions (2011)
  • Author(s): 水野健作
  • Organizer: 第84回日本生化学会大会
  • Place of Presentation: 京都
  • Year and Date: 20110921-24
• [Presentation] Multipoit measurements of fluorescence decay after photoactivation of Dronpa-actin reveal spatial distribution of G-actin concentration between lamellipodium and cell body (2011)
  • Author(s): 木内泰、永井友朗、大橋一正、水野健作
  • Organizer: 第63回日本細胞生物学会大会
  • Place of Presentation: 札幌
  • Year and Date: 20110627-0629
• [Presentation] NDR-mediated Rabin8 phosphorylation is required for ciliogenesis (2010)
  • Author(s): Chiba, S., Amagai, Y., Kanno, S., Yasui, A., Fukuda, M., and Mizuno , K.
  • Organizer: 50th American Society for Cell Biology Annual meeting
  • Place of Presentation: Pensylvenia, USA
  • Year and Date: 20101210-1217
• [Presentation] 細胞移動時の接着斑のターンオーバーにおけるSlingshotファミリーの機能解析 (2010)
  • Author(s): 高橋克宣、吉田圭一、梅原里奈、大橋一正、水野健作
  • Organizer: 第33回日本分子生物学会年会第83回日本生化学会合同大会
  • Place of Presentation: 神戸
  • Year and Date: 20101207-1210
• [Presentation] Regulation of Furry localization to spindle microtubules (2010)
  • Author(s): Ikeda, M., Chiba, S., Ohashi, K., and Mizuno, K.
  • Organizer: 第33回日本分子生物学会年会第83回日本生化学会合同大会
  • Place of Presentation: 神戸
  • Year and Date: 20101207-1210
• [Presentation] Critical roles of NDR and Furry in chromosome alignment and spindle formation (2010)
  • Author(s): Mizuno, K.
  • Organizer: 国際シンポジウム"Cell Cycle and Cell differentiation From A to Z"
  • Place of Presentation: 名古屋
  • Year and Date: 20101104-1106
• [Presentation] 細胞内アクチンモノマー濃度と刺激依存的なラメリポディア形成との強い相関関係 (2010)
  • Author(s): 永井友朗、木内泰、大橋一正、水野健作
  • Organizer: 第62回日本細胞生物学会大会ワークショップ(口演)+ ポスター
  • Place of Presentation: 大阪
  • Year and Date: 20100519-21
• [Presentation] High-throughput screening for low-molecular weight inhibitors of protein-protein interactions using BiFC probes (2009)
  • Author(s): 東海林和康、大橋一正、三瓶かおり、木内泰、水野健作
  • Organizer: 第32回日本分子生物学会年会
  • Place of Presentation: 横浜
  • Year and Date: 20091209-1212
• [Presentation] Identification of filamin A as the Slingshot-binding protein (2009)
  • Author(s): 佐上彩、太田裕作、栗田宗一、大橋一正、水野健作
  • Organizer: 第32回日本分子生物学会年会
  • Place of Presentation: 横浜
  • Year and Date: 20091209-1212
• [Presentation] Mitotic activation of NDR1 kinase and its role in chromosome alignment (2009)
  • Author(s): 池田真教、千葉秀平、大橋一正、水野健作
  • Organizer: 第32回日本分子生物学会年会
  • Place of Presentation: 横浜
  • Year and Date: 20091209-1212
• [Presentation] 癌細胞のmesenchymal-amoeboid transitionにおけるLIMキナーゼの役割 (2009)
  • Author(s): 三嶋利明、直塚萌、堀田祐司、大橋一正、水野健作
  • Organizer: 第32回日本分子生物学会年会
  • Place of Presentation: 横浜
  • Year and Date: 20091209-1201
• [Presentation] コフィリンホスファターゼSlingshotの活性制御機構と細胞遊走、細胞分裂における機能 (2009)
  • Author(s): 大橋一正、水野健作
  • Organizer: 第82回日本生化学会大会
  • Place of Presentation: 神戸
  • Year and Date: 20091021-1024
• [Presentation] NDRキナーゼの活性制御機構と染色体整列における機能 (2009)
  • Author(s): 水野健作、千葉秀平、池田真教、大橋一正
  • Organizer: 第82回日本生化学会大会
  • Place of Presentation: 神戸
  • Year and Date: 20091021-1024
• [Presentation] 生細胞内アクチンモノマー濃度変化測定のためのFDAPタイムラプスイメージング法の開発とラメリポディア形成過程におけるコフィリン、アクチンモノマーの重要性 (2009)
  • Author(s): 永井友朗、木内泰、大橋一正、水野健作
  • Organizer: 第82回日本生化学会大会
  • Place of Presentation: 神戸
  • Year and Date: 20091021-1024
• [Presentation] Wnt-PCP経路によるRhoA活性化と神経突起退縮に関与するRho-GEFの同定 (2009)
  • Author(s): 辻拓史、太田裕作、菅野祐哉、大橋一正、水野健作
  • Organizer: 第82回日本生化学会大会
  • Place of Presentation: 神戸
  • Year and Date: 20091021-1024
• [Presentation] Role of cofilin phosphorylation in BDNF-induced dendritogenesis (2009)
  • Author(s): Ohashi, K., Miyajima, K., Saito, A., Kiuchi, T., Akatsuka, J., and Mizuno, K.
  • Organizer: 第52回日本神経化学会大会(シンポジウム)
  • Place of Presentation: 伊香保
  • Year and Date: 20090621-24
• [Presentation] MST2とFurryによるNDR1の活性化は分裂期染色体の赤道面への整列に必要である (2009)
  • Author(s): 千葉秀平、池田真教、勝沼功吉、大橋一正、水野健作
  • Organizer: 第61回日本細胞生物学会大会ワークショップ(口演)
  • Place of Presentation: 名古屋
  • Year and Date: 20090602-04
• [Presentation] Dronpaを用いた細胞内G-アクチン濃度の経時測定によるコフィリンの機能解析 (2009)
  • Author(s): 永井友朗、木内泰、大橋一正、水野健作
  • Organizer: 日本生化学会東北支部例会
  • Place of Presentation: 仙台
  • Year and Date: 2009-05-09
• [Presentation] nvadopodia形成におけるコフィリンリン酸化制御の機能 (2009)
  • Author(s): 北谷佳那恵、堀田祐司、大橋一正、水野健作
  • Organizer: 日本生化学会東北支部例会
  • Place of Presentation: 仙台
  • Year and Date: 2009-05-09
• [Presentation] MST2-and Furry-mediated activation of NDR1 kinase is critical for precise alignment of mitotic chromosomes (2008)
  • Author(s): Chiba, S., Ikeda, M., Ohashi, K., and Mizuno, K.
  • Organizer: he 49th Annual Meeting of The American Society for Cell Biology
  • Place of Presentation: San Diego, Dec
  • Year and Date: 20081204-11
• [Remarks]
  • URL: http://www.biology.tohoku.ac.jp/lab-www/mizuno_lab/ohashi1234/Top.html