1998 Fiscal Year Annual Research Report

低分子量G蛋白質の活性制御機構と作用機構

Research Project

Project/Area Number	10044285
Research Category	Grant-in-Aid for international Scientific Research
Section	Joint Research .
Research Institution	Osaka University
Principal Investigator	高井義美大阪大学, 医学部, 教授 (60093514)
Co-Investigator(Kenkyū-buntansha)	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)
Keywords	低分子量G蛋白質 / Rho / Rab / Rabphilin-3A / BNI1 / SPA2 / mDia / ROCK
Research Abstract	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。

Research Products

(29 results)

All Other

All Publications (29 results)

[Publications] Ohya,T.: "Involvement of Rabphilin3 in endocytosis through interaction with Rabaptin5" J.Biol.Chem. 273・1. 316-317 (1998)
[Publications] 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)
[Publications] 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)
[Publications] 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)
[Publications] 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)
[Publications] 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)
[Publications] 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)
[Publications] 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)
[Publications] 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)
[Publications] 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)
[Publications] Ohtsuka,T.: "Nexilin-A novel actin filament-binding protein localized at cell-matrix adherens junction." J.Cell Biol.143・5. 1227-1238 (1998)
[Publications] Miki,H.: "Induction of filopodium formation by a WASP-related actin-depolymerizing protein N-WASP." Nature. 391・6662. 93-96 (1998)
[Publications] Fujita,Y.: "Tomosyn: A syntaxin-1-binding protein which forms a novel complex in the neurotransmitter release process." Neuron. 20・5. 905-915 (1998)
[Publications] Mochida,S.: "Role of the Doc2α-Munc13-1 interaction in the neurotransmitter release process." Proc.Natl.Acad.Sci.USA. 95・19. 11418-11422 (1998)
[Publications] 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)
[Publications] 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)
[Publications] Asakura,T.: "Isolation and characterization of a novel actin filament-binding protein from Saccharomyces cerevisiae." Oncogene. 16・1. 121-130 (1998)
[Publications] 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)
[Publications] Takahashi,K.: "Interaction of Radixin with Rho small G protein GDP/GTP exchange protein Db1." Oncogene. 16・25. 3279-3284 (1998)
[Publications] Burns,M.E.: "Rabphilin-3A-A multifunctional regulator of synaptic vesicle traffic." J.Gen.Physiol.111・2. 243-255 (1998)
[Publications] 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)
[Publications] 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)
[Publications] Yao,I.: "MAGUIN-A novel neuronal membrane-associated guanylate kinase-interacting protein." J.Biol.Chem.(in press). (1999)
[Publications] 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)
[Publications] Irie,M.: "Isolation and characterization of mammalian homologues of Caenorhabditis elegans lin-7-Localization at cell-cell junctions." Oncogene. (in press). (1999)
[Publications] Kodama,A.: "Involvement of Cdc42 small G protein in cell-cell adhesion,migration,and morphology of MDCK cells." Oncogene. (in press). (1999)
[Publications] 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)
[Publications] Shimizu,K.et.al.: "Methods in Molecular Biology" Human Press Inc.,Totowa,NJ, 173-183 (1998)
[Publications] Orita,S.et.al.: "Frontiers of Neural Development" Springer-Verlag Tokyo Inc.,Tokyo, 444-449 (1999)

1998 Fiscal Year Annual Research Report

低分子量G蛋白質の活性制御機構と作用機構

Principal Investigator

高井 義美 大阪大学, 医学部, 教授 (60093514)

Research Products

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

[Publications] 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)

[Publications] 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)

[Publications] 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)

[Publications] 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)

[Publications] 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)

[Publications] 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)

[Publications] 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)

[Publications] 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)

[Publications] 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)

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

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

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

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

[Publications] 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)

[Publications] 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)

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

[Publications] 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)

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

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

[Publications] 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)

[Publications] 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)

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

[Publications] 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)

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

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

[Publications] 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)

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

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

高井義美大阪大学, 医学部, 教授 (60093514)