Analysis of the mechanism of septin polymerization and depolymerization

• Project/Area Number: 18370077
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Cell biology
• Research Institution: Kyoto University
• Principal Investigator: KINOSHITA Makoto Kyoto University, 医学研究科, 講師 (30273460)
• Co-Investigator(Kenkyū-buntansha): 萩原 明 京都大学, 医学研究科, 助手 (70402849)
• Project Period (FY): 2006 – 2007
• Project Status: Completed (Fiscal Year 2007)
• Budget Amount: ¥17,620,000 (Direct Cost: ¥15,400,000、Indirect Cost: ¥2,220,000); Fiscal Year 2007: ¥9,620,000 (Direct Cost: ¥7,400,000、Indirect Cost: ¥2,220,000); Fiscal Year 2006: ¥8,000,000 (Direct Cost: ¥8,000,000)
• Keywords: biomolecule / protein / cell / tissue / brain / nervous system / cytoskeleton / reconstirution / live imaging / knockout mouse / ライブイメージング / ノックアウトマウス / 光退色後蛍光回復法 / マススペクトロメトリー / 電子顕微鏡 / 生化学

Research Abstract

セプチン系の分子動態解析を目的として、生化学的解析、超微形態解析に加えて培養神経細胞の細胞膜直下に集積したGFP-septin融合蛋白質に対するFRAP(光退色後蛍光回復)法を行った。膜蛋白質分子や細胞膜直下のアクチン分子に比べてセプチン分子のターンオーバーは緩慢であり、細胞膜直下のセプチン集合体の安定性が示唆された。この特性は細胞膜形状の保持や、膜蛋白質のスカフォールドないし拡散障壁としての役割に適したものと考えられる。RNAiでセプチンを枯渇させた細胞で関連分子の動態を調べたところ、cortical actinの動態に有意な変化が見られない条件で、syntaxin-1(膜貫通型tSNARE)やGLASTのターンオーバーが有意に促進した。したがって、細胞膜直下のセプチン系がこれらの分子のスカフォールドないし拡散障壁として局在や安定化に寄与する可能性が支持された。生化学的解析、超微形態解析に関する論文は現在執筆中である。

Research Products

• [Journal Article] Morphogenesis of polarized epithelia requires Sept2-mediated coupling of vesicle transport to polyglutamylated microtubules. (2008)
  • Author(s): Spiliotis ET, Kinoshita M, 他3名
  • Journal Title: Journal of Cell Biology 180 Pages: 295-303
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Loss of Sept4 exacerbates liver fibrosis through the dysregulation of hepatic stellate cells. (2008)
  • Author(s): Iwaisako K, Kinoshita M, 他8名
  • Journal Title: Journal of Hepatology 未定(印刷中)
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Morphogenesis of polarized epithelia requires Sept2-mediated coupling of vesicle transport to polyglutamylated microtubules (2008)
  • Author(s): Spiliotis, ET, Hunt, S., Hu, Q., Kinoshita, M., Nelson, WJ
  • Journal Title: Journal of Cell Biology 180 Pages: 295-303
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Sept4, a component of presynaptic scaffold and Lewy bodies, is required for the suppression of alpha- (2007)
  • Author(s): Ihara, Kinoshita, et. al.
  • Journal Title: Neuron 53 Pages: 519-533
  • Peer Reviewed
• [Journal Article] Connecting the dots between septins and the DNA damage checkpoint. (2007)
  • Author(s): Kinoshita, Takeda
  • Journal Title: Cell 130 Pages: 777-779
  • Peer Reviewed
• [Journal Article] Role of Septin cytoskeleton in spine morphogenesis and dendrite development in neurons. (2007)
  • Author(s): Tada, Kinoshita, Sheng, et. al.
  • Journal Title: Current Biology 17 Pages: 1752-1758
  • Peer Reviewed
• [Journal Article] Sept4, a component of presynaptic scaffold and Lewy bodies, is required for the suppression of α-synuclein neurotoxicity. (2007)
  • Author(s): Ihara M, Kinoshita M, 他11名
  • Journal Title: Neuron 53 Pages: 519-533
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Sept4, a component of presynaptic scaffold and Lewy bodies, is required for the suppression of a-synuclein neurotoxicity (2007)
  • Author(s): Ihara, M., Yamasaki, N., Hagiwara, A., Tomimoto, H., Kitano, A., Tanigaki, A., Hikawa, E., Noda, M., Takanashi, M., Mori, H., Hattori, N., Miyakawa,T., Kinoshita, M
  • Journal Title: Neuron 53 Pages: 519-533
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Isoform-specific phosphorylation of Sept5/CDCrel-1 by cyclin-dependent kinase 5 inhibits the interaction (2007)
  • Author(s): Taniguchi M, Taoka M, Itakura M, Asada A, Saito T, Kinoshita M
  • Journal Title: Journal of Biological Chemistry (Epub ahead of print) 15-Jan
• [Journal Article] Sept4, a component of presynaptic scaffold and Lewy bodies, is required for the suppression of ・-synuclein (2007)
  • Author(s): Ihara M, Yamasaki N, Hagiwara A, Tomimoto H, Kitano A, Tanigaki A
  • Journal Title: Neuron 53(3) Pages: 519-533
• [Journal Article] 蛍光偏光顕微鏡が解いたセプチン線維配向の謎 (2007)
  • Author(s): 木下 専
  • Journal Title: 蛋白質核酸酵素 52(1) Pages: 71-72
• [Journal Article] 再構成系と遺伝子破壊マウスによるセプチン系の解析 (2006)
  • Author(s): 木下 専
  • Journal Title: 細胞工学 25(7) Pages: 785-789
• [Journal Article] 細胞分裂と細胞形態形成におけるGTP結合蛋白質セプチンの役割 (2006)
  • Author(s): 木下 専
  • Journal Title: 生化学 78(8) Pages: 755-759
• [Journal Article] Loss of Sept4 exacerbates liver fibrosis through the dysregulation of hepatic stellate cells
  • Author(s): Iwaisako, K., Hatano, E., Taura, K., Nakajima, A., Tada, M., Seo, S., Tamaki, N., Ikai, I., Uemoto, S., Kinoshita, M
  • Journal Title: Journal of Hepatology (in press)
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Insight into septin functions from mouse models
  • Author(s): Kinoshita, M
  • Journal Title: In The Septins (eds. J.Pringle, and P. Hall) John Wiley & Sons (in press)
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Sept4, a component of presynaptic scaffold and Lewy bodies, is required for the suppression of alpha- (2007)
  • Author(s): Kinoshita M
  • Organizer: EMBO Workshop/The 2nd International Septin Workshop.
  • Place of Presentation: Ascona, Switzerland
  • Year and Date: 2007-05-07
• [Presentation] Sept4, a component of presynaptic scaffold and Lewy bodies, is required for the suppression of alpha-synuclein neurotoxicity. (2007)
  • Author(s): Kinoshita M
  • Organizer: EMBO Workshop/The 2nd International Septin Workshop. Keynote Lecture
  • Place of Presentation: Ascona, Switzerland
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Sept4, a component of presynaptic scaffold and Lewy bodies, is required for the suppression of alpha-synuclein neurotoxicity (2005)
  • Author(s): Kinoshita, M., Keynote, Lecture
  • Organizer: EMBO Workshop/The 2" International Septin Workshop
  • Place of Presentation: (Ascona, Switzerland)
  • Description: 「研究成果報告書概要(欧文)」より
• [Book] The Septins (eds. S.E.H. Russell, J. Pringle, and P. Hall) (2008)
  • Author(s): Kinoshita M(他11名)
  • Publisher: John Wiley & Sons(未定)
• [Book] Insight into septin functions from mouse models. In The Septions(eds. S. E. H. Russell, J. Pringle, and P. Hall) (2008)
  • Author(s): kinoshita M
  • Publisher: John Wiley & Sons(印刷中)
  • Description: 「研究成果報告書概要(和文)」より
• [Book] Insight into septin functions from mouse models. (In a monograph, "The Septins" ; eds. S.E.H. Russell, J. Pringle, and P. Hall) (2007)
  • Author(s): Kinoshita M
  • Publisher: John Wiley ＆ Sons