1998 年度実績報告書

低分子量G蛋白質の活性制御機構と作用機構

研究課題

研究課題/領域番号	10044285
研究種目	国際学術研究
応募区分	共同研究
研究機関	大阪大学
研究代表者	高井義美大阪大学, 医学部, 教授 (60093514)
研究分担者	NOVIEK Peter エール大学, 医学部, 教授 HALL Alan ロンドン大学, 理学部, 教授白瀧博通獨協医科大学, 医学総合研究所, 教授 (90249962) 佐々木卓也大阪大学, 医学部, 助教授 (40241278) 田中一馬北海道大学, 医学部, 教授 (60188290)
キーワード	低分子量G蛋白質 / Rho / Rab / Rabphilin-3A / BNI1 / SPA2 / mDia / ROCK
研究概要	現在、低分子量G蛋白質は、種々の重要な細胞機能を制御していることが明らかになっており、その活性化機構と作用機構を物質的に解明することが、医学・生物学領域において重要な課題となっている。本年度の本研究において、私共は、RhoとRab低分子量G蛋白質の活性化機構と作用機構について解析を行い、以下の成果を得た。 (1) Rab3Aの活性化機構:私共が見い出したRab3Aの活性制御蛋白質Rab3 GEPとRab3 GAPが、Rab3Aと共に神経終末に局在し、Rab3Aの活性を制御することによってCa^<2+>依存性分泌を制御していることを明らかにした。 (2) Rab3Aの作用機構:ヤリイカの巨大シナプスの系により、Rab3Aの標的蛋白質Rabphllin-3Aが実際に神経伝達物質の放出を制御していることを明らかにした。 (3) Rhoの活性化機構: MDCK細胞のTPAによる細胞遊走の際には、Rhoが時間的・空間的にOn/Offとなり、ストレスファイバーや接着斑の形成を介して細胞運動を制御していることを明らかにした。 (4) Rhoの作用機構:私共は、出芽酵母において、Rhoの標的蛋白質BNI1がアクチンの重合過程を制御することを明らかにしているが、本研究では、このBNIlを細胞膜へ局在させる蛋白質としてSPA2を見い出し、さらに、SPA2に結合する因子が細胞質分裂に関与するセプチンであることを見い出した。また、動物細胞においては、BNIlのカウンターパートであるmDiaと今一つのRhoの標的蛋白質であるROCKの機能を検討し、Rhoによるアクチン細胞骨格の再編成には、両者の協調的な作用が重要であることを明らかにした。本年度は、海外研究分担者との共同研究という形で論文は発表できなかったが、Novick博士およびHall博士との研究打ち合わせやLacal博士およびBar-Sagi博士による研究技術指導により、本年度の研究は上記のように予想以上に進展し、当初の目的はほぼ達成できた。

研究成果

(29件)

すべてその他

すべて文献書誌 (29件)

[文献書誌] Ohya,T.: "Involvement of Rabphilin3 in endocytosis through interaction with Rabaptin5" J.Biol.Chem. 273・1. 316-317 (1998)
[文献書誌] Satoh,A.: "Neurabin-II-Spinophilin-An actin filament-binding protein with one PDZ domain localized at cadherin-based cell-cell adhesion sites." J.Biol.Chem.273・6. 3470-3475 (1998)
[文献書誌] Shimizu,K.: "Complex formation of SMAP/KAP3, a KIF3A/B ATPase motor-associated protein,with a human chromosome-associated polypeptide." J.Biol.Chem.273・12. 6591-6594 (1998)
[文献書誌] Obaishi,H.: "Frabin,a novel FGD1-related actin filament-binding protein capable of changing cell shape and activating c-Jun N-terminal kinase." J.Biol.Chem.273・30. 18697-18700 (1998)
[文献書誌] Hirao,K.: "A novel multiple PDZ domain-containing molecule interacting with N-methyl-D-aspartate receptors and neuronal cell adhesion proteins." J.Biol.Chem.273・33. 21105-21110 (1998)
[文献書誌] Nagano,F.: "Molecular cloning and characterization of the noncatalytic subunit of the Rab3 subfamily-specific GTPase-activating protein." J.Biol.Chem.273・38. 24781-24785 (1998)
[文献書誌] Deguchi,M.: "BEGAIN (Brain-enriched guanylate kinase-associated protein),a novel neuronal PSD-95/SAP90-binding protein." J.Biol.Chem.273・41. 26269-26272 (1998)
[文献書誌] Kamei,T.: "Interaction of Bnrlp with a novel Src homology 3 domain-containing Hoflp-Inplication in cytokinesis in Saccharomyces cerevisiae." J.Biol.Chem.273・43. 28341-28345 (1998)
[文献書誌] Nagano,F.: "Interaction of Doc2 with tctex-1,a light chain of cytoplasmic dynein-Implication in dynein-dependent vesicle transport." J.Biol.Chem.273・46. 30065-30068 (1998)
[文献書誌] Oishi,H.: "Localization of the Rab3 small G protein regulators in nerve terminals and their involvement in Ca^<2+>-dependent exocytosis." J.Biol.Chem.273・51. 34580-34585 (1998)
[文献書誌] Ohtsuka,T.: "Nexilin-A novel actin filament-binding protein localized at cell-matrix adherens junction." J.Cell Biol.143・5. 1227-1238 (1998)
[文献書誌] Miki,H.: "Induction of filopodium formation by a WASP-related actin-depolymerizing protein N-WASP." Nature. 391・6662. 93-96 (1998)
[文献書誌] Fujita,Y.: "Tomosyn: A syntaxin-1-binding protein which forms a novel complex in the neurotransmitter release process." Neuron. 20・5. 905-915 (1998)
[文献書誌] Mochida,S.: "Role of the Doc2α-Munc13-1 interaction in the neurotransmitter release process." Proc.Natl.Acad.Sci.USA. 95・19. 11418-11422 (1998)
[文献書誌] Fujiwara,T.: "Rho1p-Bni1p-Spa2p interactions-Implication in localization of Bni1p at the bud site and regulation of the actin cytoskeleton in Saccharomyces cerevisiae." Mol.Biol.Cell. 9・5. 1221-1223 (1998)
[文献書誌] Imamura,H.: "Rho and Rab small G proteins coordinately reorganize stress fibers and focal adhesions in MDCK cells." Mol.Biol.Cell. 9・9. 2561-2575 (1998)
[文献書誌] Asakura,T.: "Isolation and characterization of a novel actin filament-binding protein from Saccharomyces cerevisiae." Oncogene. 16・1. 121-130 (1998)
[文献書誌] Umikawa,M.: "Interaction of Rho1p target Bni1p and F-actin-binding elongation factor 1α-Implication in Rho1p-regulated reorganization of the actin cytoskeleton in Saccharomyces cerevisiae." Oncogene. 16・15. 2011-2016 (1998)
[文献書誌] Takahashi,K.: "Interaction of Radixin with Rho small G protein GDP/GTP exchange protein Db1." Oncogene. 16・25. 3279-3284 (1998)
[文献書誌] Burns,M.E.: "Rabphilin-3A-A multifunctional regulator of synaptic vesicle traffic." J.Gen.Physiol.111・2. 243-255 (1998)
[文献書誌] Tanaka,K.: "Control of reorganization of the actin cytoskeleton by Rho family small GTP-binding proteins in yeast." Curr.Opin.Cell Biol.10・1. 112-116 (1998)
[文献書誌] Sasaki,T.: "The Rho small G protein family-Rho GDI system as a temporal and spatial determinant for cytoskeletal control." Biochem.Biophys.Res.Commun.245・3. 641-645 (1998)
[文献書誌] Yao,I.: "MAGUIN-A novel neuronal membrane-associated guanylate kinase-interacting protein." J.Biol.Chem.(in press). (1999)
[文献書誌] Sakisaka,T.: "Different behavior of 1-afadin and and neurabin-II during the formation and destruction of cell-cell adherens junction." Oncogene(in press). (in press). (1999)
[文献書誌] Irie,M.: "Isolation and characterization of mammalian homologues of Caenorhabditis elegans lin-7-Localization at cell-cell junctions." Oncogene. (in press). (1999)
[文献書誌] Kodama,A.: "Involvement of Cdc42 small G protein in cell-cell adhesion,migration,and morphology of MDCK cells." Oncogene. (in press). (1999)
[文献書誌] Mandai K.: "Ponsin-SH3P12: An 1-afadin-and vinculin-binding protein localized at cell-cell and cell-matrix adherens junctions." J.Cell Biol.(in press). (1999)
[文献書誌] Shimizu,K.et.al.: "Methods in Molecular Biology" Human Press Inc.,Totowa,NJ, 173-183 (1998)
[文献書誌] Orita,S.et.al.: "Frontiers of Neural Development" Springer-Verlag Tokyo Inc.,Tokyo, 444-449 (1999)

1998 年度 実績報告書

低分子量G蛋白質の活性制御機構と作用機構

研究代表者

高井 義美 大阪大学, 医学部, 教授 (60093514)

研究成果

[文献書誌] Ohya,T.: "Involvement of Rabphilin3 in endocytosis through interaction with Rabaptin5" J.Biol.Chem. 273・1. 316-317 (1998)

[文献書誌] Satoh,A.: "Neurabin-II-Spinophilin-An actin filament-binding protein with one PDZ domain localized at cadherin-based cell-cell adhesion sites." J.Biol.Chem.273・6. 3470-3475 (1998)

[文献書誌] Shimizu,K.: "Complex formation of SMAP/KAP3, a KIF3A/B ATPase motor-associated protein,with a human chromosome-associated polypeptide." J.Biol.Chem.273・12. 6591-6594 (1998)

[文献書誌] Obaishi,H.: "Frabin,a novel FGD1-related actin filament-binding protein capable of changing cell shape and activating c-Jun N-terminal kinase." J.Biol.Chem.273・30. 18697-18700 (1998)

[文献書誌] Hirao,K.: "A novel multiple PDZ domain-containing molecule interacting with N-methyl-D-aspartate receptors and neuronal cell adhesion proteins." J.Biol.Chem.273・33. 21105-21110 (1998)

[文献書誌] Nagano,F.: "Molecular cloning and characterization of the noncatalytic subunit of the Rab3 subfamily-specific GTPase-activating protein." J.Biol.Chem.273・38. 24781-24785 (1998)

[文献書誌] Deguchi,M.: "BEGAIN (Brain-enriched guanylate kinase-associated protein),a novel neuronal PSD-95/SAP90-binding protein." J.Biol.Chem.273・41. 26269-26272 (1998)

[文献書誌] Kamei,T.: "Interaction of Bnrlp with a novel Src homology 3 domain-containing Hoflp-Inplication in cytokinesis in Saccharomyces cerevisiae." J.Biol.Chem.273・43. 28341-28345 (1998)

[文献書誌] Nagano,F.: "Interaction of Doc2 with tctex-1,a light chain of cytoplasmic dynein-Implication in dynein-dependent vesicle transport." J.Biol.Chem.273・46. 30065-30068 (1998)

[文献書誌] Oishi,H.: "Localization of the Rab3 small G protein regulators in nerve terminals and their involvement in Ca^<2+>-dependent exocytosis." J.Biol.Chem.273・51. 34580-34585 (1998)

[文献書誌] Ohtsuka,T.: "Nexilin-A novel actin filament-binding protein localized at cell-matrix adherens junction." J.Cell Biol.143・5. 1227-1238 (1998)

[文献書誌] Miki,H.: "Induction of filopodium formation by a WASP-related actin-depolymerizing protein N-WASP." Nature. 391・6662. 93-96 (1998)

[文献書誌] Fujita,Y.: "Tomosyn: A syntaxin-1-binding protein which forms a novel complex in the neurotransmitter release process." Neuron. 20・5. 905-915 (1998)

[文献書誌] Mochida,S.: "Role of the Doc2α-Munc13-1 interaction in the neurotransmitter release process." Proc.Natl.Acad.Sci.USA. 95・19. 11418-11422 (1998)

[文献書誌] Fujiwara,T.: "Rho1p-Bni1p-Spa2p interactions-Implication in localization of Bni1p at the bud site and regulation of the actin cytoskeleton in Saccharomyces cerevisiae." Mol.Biol.Cell. 9・5. 1221-1223 (1998)

[文献書誌] Imamura,H.: "Rho and Rab small G proteins coordinately reorganize stress fibers and focal adhesions in MDCK cells." Mol.Biol.Cell. 9・9. 2561-2575 (1998)

[文献書誌] Asakura,T.: "Isolation and characterization of a novel actin filament-binding protein from Saccharomyces cerevisiae." Oncogene. 16・1. 121-130 (1998)

[文献書誌] Umikawa,M.: "Interaction of Rho1p target Bni1p and F-actin-binding elongation factor 1α-Implication in Rho1p-regulated reorganization of the actin cytoskeleton in Saccharomyces cerevisiae." Oncogene. 16・15. 2011-2016 (1998)

[文献書誌] Takahashi,K.: "Interaction of Radixin with Rho small G protein GDP/GTP exchange protein Db1." Oncogene. 16・25. 3279-3284 (1998)

[文献書誌] Burns,M.E.: "Rabphilin-3A-A multifunctional regulator of synaptic vesicle traffic." J.Gen.Physiol.111・2. 243-255 (1998)

[文献書誌] Tanaka,K.: "Control of reorganization of the actin cytoskeleton by Rho family small GTP-binding proteins in yeast." Curr.Opin.Cell Biol.10・1. 112-116 (1998)

[文献書誌] Sasaki,T.: "The Rho small G protein family-Rho GDI system as a temporal and spatial determinant for cytoskeletal control." Biochem.Biophys.Res.Commun.245・3. 641-645 (1998)

[文献書誌] Yao,I.: "MAGUIN-A novel neuronal membrane-associated guanylate kinase-interacting protein." J.Biol.Chem.(in press). (1999)

[文献書誌] Sakisaka,T.: "Different behavior of 1-afadin and and neurabin-II during the formation and destruction of cell-cell adherens junction." Oncogene(in press). (in press). (1999)

[文献書誌] Irie,M.: "Isolation and characterization of mammalian homologues of Caenorhabditis elegans lin-7-Localization at cell-cell junctions." Oncogene. (in press). (1999)

[文献書誌] Kodama,A.: "Involvement of Cdc42 small G protein in cell-cell adhesion,migration,and morphology of MDCK cells." Oncogene. (in press). (1999)

[文献書誌] Mandai K.: "Ponsin-SH3P12: An 1-afadin-and vinculin-binding protein localized at cell-cell and cell-matrix adherens junctions." J.Cell Biol.(in press). (1999)

[文献書誌] Shimizu,K.et.al.: "Methods in Molecular Biology" Human Press Inc.,Totowa,NJ, 173-183 (1998)

[文献書誌] Orita,S.et.al.: "Frontiers of Neural Development" Springer-Verlag Tokyo Inc.,Tokyo, 444-449 (1999)

1998 年度実績報告書

高井義美大阪大学, 医学部, 教授 (60093514)