1996 Fiscal Year Annual Research Report

細胞の増殖・分化のシグナル伝達機構

Research Project

Project/Area Number	06283221
Research Institution	Osaka University
Principal Investigator	高井義美大阪大学, 医学部, 教授 (60093514)
Co-Investigator(Kenkyū-buntansha)	佐々木卓也大阪大学, 医学部, 助手 (40241278) 田中一馬大阪大学, 医学部, 助教授 (60188290)
Keywords	細胞骨格 / Rho / ERM / Rho GDI / BNI1 / CD44 / アクチン
Research Abstract	細胞が、がん化して浸潤能や転移能を獲得する過程において、細胞の形態や運動性、極性、接着性、分泌性等々の異常が重要な役割を果たしているが、これらの細胞機能は主としてアクチン系の細胞骨格により制御されている。一方、Rho低分子量G蛋白質が、私共を含めたいくつかのグループによって、アクチン細胞骨格の再編成を介して、上述した細胞機能を制御していることが明らかにされつつあり、Rhoを介するシグナル伝達経路の異常が、がん細胞の浸潤や転移において重要な役割を果たしている可能性が高い。本研究では、Rhoの活性化機構と作用機構に焦点をしぼって解析を行い、以下の成果を得た。私共は、アクチン繊維を細胞膜へAnchoringするERM(Ezrin,Radixin,Moesin)-CD44系が、Rhoの作用点として機能していることを明らかにしているが、このERMが、Rho-Rho GDI複合体に直接作用し、RhoをRho GDIから解離させて活性化反応を開始させる作用も有していることを明らかにした。一方、活性化されたRhoは、BNI1様標的蛋白質に作用し、アクチン結合蛋白質であるプロフィリンを介してアクチンの重合過程を制御すると共に、何らかの標的蛋白質を介して、ERMの細胞膜への移行過程、すなわちCD44との会合を制御することにより、アクチン細胞骨格の再編成を統合的に制御していることを明らかにした。以上の成果を得たことにより、本年度の研究は順調に進んだと考えている。

Research Products

(30 results)

All Other

All Publications (30 results)

[Publications] Ohtsuka,T.: "Activation of brain B-Raf protein kinase by RaplB small GTP-binding protein" J.Biol.Chem.271. 1258-1261 (1996)
[Publications] Orita,S.: "Doc2 enhances Ca^<2+>-dependent exocytosis from PC12 cells" J.Biol.Chem.271. 7257-7260 (1996)
[Publications] Fujita,Y.: "phosphorylation of Munc-18/n-Secl/rbSecl by protein kinase C-Its implication in regulating the interaction of Munc-18/n-Secl/rbSecl with Syntaxin" J.Biol.Chem.271. 7265-7268 (1996)
[Publications] Kuroda,S.: "Different effects of various phospholipids on Ki-Ras-,Ha-Ras,and RaplB-induced B-Raf activation" J.Biol.Chem.271. 14680-14683 (1996)
[Publications] Simizu,K.: "SMAP,an Smg GDS-associating protein having Arm repeats and phosphorylated by Src tyrosine kinase" J.Biol.Chem.271. 27013-27017 (1996)
[Publications] Kato,M.: "Physical and functional interaction of Rabphilin-3A with α-Actinin." J.Biol.Chem. 271. 31775-31778 (1996)
[Publications] Hirano,H.: "ROM7/BEM4 encodes a novel protein that interacts with the Rholp small GTP-binding protein in Saccharomyces cerevisiae" Mol.Cell.Biol.16. 4396-4403 (1996)
[Publications] Hirao,M.: "Regulation mechanism of ERM(Ezrin/Radixin/Moesin)protein/plasma membrane association - Possible involvement of phosphatidylinositol turnover and Rho-dependent signaling pathway" J.Cell.Biol.135. 37-51 (1996)
[Publications] Masumoto,N.: "Involvement of Rabphilin-3A in cortical granule exocytosis in mouse eggs" J.Cell.Biol.135. 1741-1747 (1996)
[Publications] Drgonova,J.: "Rholp,a yeast protein at the interface between cell polarization and morphogenesis" Science. 272. 277-279 (1996)
[Publications] Ozaki,K.: "Romlp and Rom2p are GDP/GTP exchange proteins(GEPs) for the Rholp small GTP binding protein in Saccharomyces cerevisiae" EMBO J.15. 2196-2207 (1996)
[Publications] Kohno,H.: "Bnilp implicated in cytoskeletal control is a putative target of Rholp small GTP binding protein in Saccharomyces cerevisiae" EMBO J.15. 6060-6068 (1996)
[Publications] Matsuda,S.: "A membrane-associated GDP/GTP exchange protein specific for Rho small GTP-binding protein - Partial purification and characterization from rat brain" Oncogene. 12. 915-920 (1996)
[Publications] Taniguchi,T.: "Cholecystokinin-B/gastrin receptors mediate rapid formation of actin stress fibers" Oncogene. 12. 1357-1360 (1996)
[Publications] Komuro,R.: "Involvement of rabphilin-3A in Ca^<2+>-dependent exocytosis from PC12 cells" Biochem.Biophys.Res.Commun.219. 435-440 (1996)
[Publications] Hotta,K.: "Interaction of the Rho family small G proteins with kinectin,an anchoring protein of kinesin motor" Biochem.Biophys.Res.Commun.225. 69-74 (1996)
[Publications] Senbonmatsu,T.: "Interaction of Rabphilin3 with synaptic vesicles through multiple regions" Biochem.Biophys.Res.Commun.228. 567-572 (1996)
[Publications] Oishi,H.: "Interaction of both the C2A and C2B domains of rabphilin3 with Ca^<2+> and pospholipid" Biochem.Biophys.Res.Commun.229. 498-503 (1996)
[Publications] Hori,Y.: "Possible involvement of Rabll p24,a ras-like small GTP-binding protein,in intracellular vesicular transport of isolated pancreatic acini" Digestive Diseases and Sciences. 41. 133-138 (1996)
[Publications] Regazzi,R.: "Expression,localization and functional role of small GTPases of the Rab3 family in insulinsecreting cells" J.Cell Sci.109. 2265-2273 (1996)
[Publications] Komuro,R.: "Involvement of Rho and Rac small G proteins and Rho GDI in Ca^<2+>-dependent exocytosis from PC12 cells" Genes to Cells. 1. 943-951 (1996)
[Publications] Takai,Y.: "Rab3A small GTP-binding protein in Ca^<2+>-dependent exocytosis" Genes to Cells. 1. 615-632 (1996)
[Publications] Wada,M.: "Isolation and characterization of a GDP/GTP exchange protein specific for the Rab3 subfamily small G proteins" J.Biol.Chem.272. 3875-3878 (1997)
[Publications] Fukui,K.: "Isolation and characterization of a GTPase activating protein specific for the Rab3 subfamily of small G proteins" J.Biol.Chem.272. 4655-4658 (1997)
[Publications] Takeuchi,M.: "SAPAPs-A family of PSD-95/SAP90- associated proteins localized at postsynaptic density" J.Biol.Chem.(in press). (1997)
[Publications] Imamura,H.: "Bnilp and Bnrlp - Downstream targets of the Rho family small G proteins which interact with profilin and regulate actin cytoskeleton in Saccharomyces cerevisiae" EMBO J.(in press). (1997)
[Publications] Kotani,H.: "Rho regulates association of both the ERM family and vinculin with the plasma membrane in MDCK cells" Oncogene. (in press). (1997)
[Publications] Naito,A.: "Molecular cloning of mouse Doc2α and distribution of its mRNA in adult mouse brain" Mol.Brain.Res.(in press). (1997)
[Publications] Jin-no,Y.: "A novel function of the C-terminal lipid moieties of Rab3A small G protein implicated in Ca2+-dependent exocytosis -Inhibition of interaction with GTP and reduction of this inhibition by phospholipid" Genes to Cells. (in press). (1997)
[Publications] Koch,G.A.: "Association of the Rho family small GPT-binding proteins with Rho GDP dissociation inhibitor(Rho GDI)in Saccharomyces cerevisiae" Oncogene. (in press). (1997)

1996 Fiscal Year Annual Research Report

細胞の増殖・分化のシグナル伝達機構

Principal Investigator

高井 義美 大阪大学, 医学部, 教授 (60093514)

Research Products

[Publications] Ohtsuka,T.: "Activation of brain B-Raf protein kinase by RaplB small GTP-binding protein" J.Biol.Chem.271. 1258-1261 (1996)

[Publications] Orita,S.: "Doc2 enhances Ca^<2+>-dependent exocytosis from PC12 cells" J.Biol.Chem.271. 7257-7260 (1996)

[Publications] Fujita,Y.: "phosphorylation of Munc-18/n-Secl/rbSecl by protein kinase C-Its implication in regulating the interaction of Munc-18/n-Secl/rbSecl with Syntaxin" J.Biol.Chem.271. 7265-7268 (1996)

[Publications] Kuroda,S.: "Different effects of various phospholipids on Ki-Ras-,Ha-Ras,and RaplB-induced B-Raf activation" J.Biol.Chem.271. 14680-14683 (1996)

[Publications] Simizu,K.: "SMAP,an Smg GDS-associating protein having Arm repeats and phosphorylated by Src tyrosine kinase" J.Biol.Chem.271. 27013-27017 (1996)

[Publications] Kato,M.: "Physical and functional interaction of Rabphilin-3A with α-Actinin." J.Biol.Chem. 271. 31775-31778 (1996)

[Publications] Hirano,H.: "ROM7/BEM4 encodes a novel protein that interacts with the Rholp small GTP-binding protein in Saccharomyces cerevisiae" Mol.Cell.Biol.16. 4396-4403 (1996)

[Publications] Hirao,M.: "Regulation mechanism of ERM(Ezrin/Radixin/Moesin)protein/plasma membrane association - Possible involvement of phosphatidylinositol turnover and Rho-dependent signaling pathway" J.Cell.Biol.135. 37-51 (1996)

[Publications] Masumoto,N.: "Involvement of Rabphilin-3A in cortical granule exocytosis in mouse eggs" J.Cell.Biol.135. 1741-1747 (1996)

[Publications] Drgonova,J.: "Rholp,a yeast protein at the interface between cell polarization and morphogenesis" Science. 272. 277-279 (1996)

[Publications] Ozaki,K.: "Romlp and Rom2p are GDP/GTP exchange proteins(GEPs) for the Rholp small GTP binding protein in Saccharomyces cerevisiae" EMBO J.15. 2196-2207 (1996)

[Publications] Kohno,H.: "Bnilp implicated in cytoskeletal control is a putative target of Rholp small GTP binding protein in Saccharomyces cerevisiae" EMBO J.15. 6060-6068 (1996)

[Publications] Matsuda,S.: "A membrane-associated GDP/GTP exchange protein specific for Rho small GTP-binding protein - Partial purification and characterization from rat brain" Oncogene. 12. 915-920 (1996)

[Publications] Taniguchi,T.: "Cholecystokinin-B/gastrin receptors mediate rapid formation of actin stress fibers" Oncogene. 12. 1357-1360 (1996)

[Publications] Komuro,R.: "Involvement of rabphilin-3A in Ca^<2+>-dependent exocytosis from PC12 cells" Biochem.Biophys.Res.Commun.219. 435-440 (1996)

[Publications] Hotta,K.: "Interaction of the Rho family small G proteins with kinectin,an anchoring protein of kinesin motor" Biochem.Biophys.Res.Commun.225. 69-74 (1996)

[Publications] Senbonmatsu,T.: "Interaction of Rabphilin3 with synaptic vesicles through multiple regions" Biochem.Biophys.Res.Commun.228. 567-572 (1996)

[Publications] Oishi,H.: "Interaction of both the C2A and C2B domains of rabphilin3 with Ca^<2+> and pospholipid" Biochem.Biophys.Res.Commun.229. 498-503 (1996)

[Publications] Hori,Y.: "Possible involvement of Rabll p24,a ras-like small GTP-binding protein,in intracellular vesicular transport of isolated pancreatic acini" Digestive Diseases and Sciences. 41. 133-138 (1996)

[Publications] Regazzi,R.: "Expression,localization and functional role of small GTPases of the Rab3 family in insulinsecreting cells" J.Cell Sci.109. 2265-2273 (1996)

[Publications] Komuro,R.: "Involvement of Rho and Rac small G proteins and Rho GDI in Ca^<2+>-dependent exocytosis from PC12 cells" Genes to Cells. 1. 943-951 (1996)

[Publications] Takai,Y.: "Rab3A small GTP-binding protein in Ca^<2+>-dependent exocytosis" Genes to Cells. 1. 615-632 (1996)

[Publications] Wada,M.: "Isolation and characterization of a GDP/GTP exchange protein specific for the Rab3 subfamily small G proteins" J.Biol.Chem.272. 3875-3878 (1997)

[Publications] Fukui,K.: "Isolation and characterization of a GTPase activating protein specific for the Rab3 subfamily of small G proteins" J.Biol.Chem.272. 4655-4658 (1997)

[Publications] Takeuchi,M.: "SAPAPs-A family of PSD-95/SAP90- associated proteins localized at postsynaptic density" J.Biol.Chem.(in press). (1997)

[Publications] Imamura,H.: "Bnilp and Bnrlp - Downstream targets of the Rho family small G proteins which interact with profilin and regulate actin cytoskeleton in Saccharomyces cerevisiae" EMBO J.(in press). (1997)

[Publications] Kotani,H.: "Rho regulates association of both the ERM family and vinculin with the plasma membrane in MDCK cells" Oncogene. (in press). (1997)

[Publications] Naito,A.: "Molecular cloning of mouse Doc2α and distribution of its mRNA in adult mouse brain" Mol.Brain.Res.(in press). (1997)

[Publications] Jin-no,Y.: "A novel function of the C-terminal lipid moieties of Rab3A small G protein implicated in Ca2+-dependent exocytosis -Inhibition of interaction with GTP and reduction of this inhibition by phospholipid" Genes to Cells. (in press). (1997)

[Publications] Koch,G.A.: "Association of the Rho family small GPT-binding proteins with Rho GDP dissociation inhibitor(Rho GDI)in Saccharomyces cerevisiae" Oncogene. (in press). (1997)

高井義美大阪大学, 医学部, 教授 (60093514)