多細胞生物の可塑性を支える小胞輸送の制御機構

• Project/Area Number: 15079207
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: The University of Tokushima
• Principal Investigator: 佐々木 卓也 The University of Tokushima, 大学院・ヘルスバイオサイエンス研究部, 教授 (40241278)
• Co-Investigator(Kenkyū-buntansha): 西村 範行 徳島大学, 大学院・ヘルスバイオサイエンス研究部, 准教授 (00322719)
• Project Period (FY): 2003 – 2007
• Project Status: Completed (Fiscal Year 2007)
• Budget Amount: ¥105,200,000 (Direct Cost: ¥105,200,000); Fiscal Year 2007: ¥18,400,000 (Direct Cost: ¥18,400,000); Fiscal Year 2006: ¥19,600,000 (Direct Cost: ¥19,600,000); Fiscal Year 2005: ¥19,600,000 (Direct Cost: ¥19,600,000); Fiscal Year 2004: ¥23,800,000 (Direct Cost: ¥23,800,000); Fiscal Year 2003: ¥23,800,000 (Direct Cost: ¥23,800,000)
• Keywords: 小胞輸送 / 可塑性 / 神経伝達 / 細胞接着 / Rabファミリー低分子量GTPase / Rabconnection-3 / Rab13 / JRAB / 低分子量CTPase / Rab3 GAP / 細胞内小胞輸送 / Rabファミリー低分子量G蛋白質 / Rab3A / タイトジャンクション / 接着分子 / claudin / occludin / Rab / 高次神経機能 / 組織・器官形成 / 細胞間接着分子

Research Abstract

本研究では、Rabファミリー低分子量G蛋白質を中心とした細胞内小胞輸送の制御分子群に焦点を絞って解析し、多細胞生物の可塑性を支える小胞輸送の制御機構について解析を行った。情報の可塑性については、Rab3の活性制御蛋白質であるRab3GEPに直接結合し、Rab3GAPに間接的に結合するシナプス小胞蛋白質Rabconnection-3についてマウス個体と海馬の初代神経培養細胞を用いた解析を行い、本蛋白質がシナプス形成に関与している可能性を示す結果を得ている。一方、形態の可塑性については、これまでに私どもはタイトジャンクション(TJ)の細胞間接着分子であるoccludinのリサイクリングを制御するRab13-JRAB系を見出しているが、本年度はRab13-JRAB系がTJの今ひとつの接着分子claudinの輸送にも関与することを示した。さらに、JRABはRab13以外にRab8とも結合し、Rab8-JRAB系は、アドヘレンスジャンクションの接着分子E-cadherinの細胞膜への輸送に関与することを明らかにした。一方、これまでにJRABがアクチン細胞骨格系に沿って局在することを示していたが、本年度の本研究で、JRABがアクチン結合蛋白質のひとつであるアクチニン4と直接結合することを示した。また、JRABはアクチニン4と結合することによって細胞膜にリクルートされて接着分子の輸送に機能すること、その結合はRab13により促進されることを示した。このように、本年度の本研究は順調に進展し、当初の目的はほぼ達成できた。

Research Products

• [Journal Article] The interaction of JRAB/MICAL-L2 with Rab8 and Rab13 coordinates the assembly of tight junctions and adherens junctions. (2008)
  • Author(s): Yamamura, R.
  • Journal Title: Mol. Biol. Cell 19 Pages: 971-983
  • Peer Reviewed
• [Journal Article] Doc2a and Munc13-4 regulate Ca^<2+>-dependent secretory lysosome exocytosis in (2008)
  • Author(s): Higashio, H.
  • Journal Title: J.immunol. 180 Pages: 4774-4784
  • Peer Reviewed
• [Journal Article] Involvement of Rab13 and JRAB/MICAL-L2 in epithelial cell scattering. (2008)
  • Author(s): Kanda, I.
  • Journal Title: Oncogene 27 Pages: 1687-1695
  • Peer Reviewed
• [Journal Article] Involvement of Actinin-4 in the recruitment of JRAB/MICAL-L2 to cell-cell Junctions and the formation of functional tight junctions. (2008)
  • Author(s): Nakatsuji, H.
  • Journal Title: Mol. Cell. Biol. (in press)
  • Peer Reviewed
• [Journal Article] Interaction of Rab3B with microtubule-binding protein Gas8 in NIH 3T3 cells. (2008)
  • Author(s): Nishimura, H.
  • Journal Title: Arch, Biochem. Biophys. (in press)
  • Peer Reviewed
• [Journal Article] Rab family small G proteins in regulation of epithelial apical junctions. (2008)
  • Author(s): Nishimura, H.
  • Journal Title: Front. Biosci. (in press)
  • Peer Reviewed
• [Journal Article] Cell-surface biotinylation to study endocytosis an d recycling of occludin. (2008)
  • Author(s): Nishimura, H.
  • Journal Title: Methods Mol. Biol. 440 Pages: 89-96
• [Journal Article] Regulation of epithelial cell adhesion and repulsion : role of endocytic recyching. (2008)
  • Author(s): Nishimura, H.
  • Journal Title: J. Med. Invest. 55 Pages: 9-16
  • Peer Reviewed
• [Journal Article] Analysis on the emerging role of Rab3 GTP ase-activating protein in Warburg Micro and Martsolf syndrome. (2008)
  • Author(s): Sakane, A.
  • Journal Title: Methods Enzymol. (in press)
• [Journal Article] Identification and characterization of JRAB/MICAL-L2, a junctional Rab (2008)
  • Author(s): Nishimura, N.
  • Journal Title: Methods Enzymol. (in press)
• [Journal Article] Involvement of Rabring7 in EGF receptor degradation as an E3 ligase. (2007)
  • Author(s): Sakane, A.
  • Journal Title: Biochem. Biophys. Res. Commun 357 Pages: 1058-1064
  • Peer Reviewed
• [Journal Article] Involvement of Rabring7 in EGF receptor degradation as an E3 ligase (2007)
  • Author(s): Sakane, A.
  • Journal Title: Biochem. Biophys. Res. Commun. (in press)
• [Journal Article] Cell-surface biothinylation to study endocytosis and recycling of occludin (2007)
  • Author(s): Nishimura, N.
  • Journal Title: Methods Mol. Biol. (in press)
• [Journal Article] JRAB/MICAL-L2 is a junctional Rab13-binding protein mediating the endocytic recycling of occludi (2006)
  • Author(s): Terai, T.
  • Journal Title: Mol. Biol. Cell 17 Pages: 2465-2475
• [Journal Article] Rab3 GTPase-activating proteinz regulates synaptic transmission and plasticity through the inactivation of Rab3 (2006)
  • Author(s): Sakane, A.
  • Journal Title: Proc. Natl. Acad. Sci. USA 103 Pages: 10029-10034
• [Journal Article] JRAB/MICAL-L2 is a junctional Rab13-binding protein mediating the endocytic recycling of occludin (2006)
  • Author(s): Terai, T.
  • Journal Title: Mol.Biol.Cell in press
• [Journal Article] Rab13 mediates the continuous endocytic recycling of occluding to the surface (2005)
  • Author(s): Morimoto, S.
  • Journal Title: J.Biol.Chem. 280 Pages: 2220-2228
• [Journal Article] Rabring7 : A target protein for Rab7 small G protein (2005)
  • Author(s): Mizuno, K.
  • Journal Title: Methods Enzymol. 403 Pages: 687-696
• [Journal Article] Rab13 mediates the continuous endocytic recycling of occludin to the cell surface (2005)
  • Author(s): Morimoto, S.
  • Journal Title: J.Biol.Chem. 280 Pages: 2220-2228
• [Journal Article] Involvement of Rab27 in antigen-induced histamine release from rat basophilic leukemia 2H3 cells (2004)
  • Author(s): Goishi, K.
  • Journal Title: Biochem.Biophys.Res.Commun. 324 Pages: 294-301
• [Journal Article] Identification and characterization of Noc2 as a potential Rab3B effector protein in epithelial cells (2004)
  • Author(s): Manabe, S.
  • Journal Title: Biochem.Biophys.Res.Commun. 316 Pages: 218-225
• [Presentation] Functions of Rab family small G proteins in regulated exocytosis. (2007)
  • Author(s): Takuya Sasaki
  • Organizer: International Symposium on Membrane Traffic
  • Place of Presentation: Awaji
  • Year and Date: 2007-11-29
• [Publications] Mizuno, K.: "Rabring7,a novel Rab7 target protein with a RING finger motif"Mol.Biol.Cell. 14. 3741-3752 (2003)
• [Publications] Yamamoto, Y.: "Distinct roles of Rab3B and Rab13 in the polarized transport of apical, basolateral, and tight junctional membrane proteins to the plasma membrane"Biochem.Biophys.Res.Commun.. 308. 270-275 (2003)
• [Publications] Nakahara, H.: "Involvement of Cdc42 and Rac small G proteins in invadopodia formation of RPMI7951 cells"Genes Cells. 8. 1019-1027 (2003)
• [Publications] Manabe, S.: "Identificatin and characterization of Noc2 as a potential Rab3B effector protein in epithelial cells"Biochem.Biophys.Res.Commun.. 316. 218-225 (2004)