1992 Fiscal Year Annual Research Report

低分子量GTP結合蛋白質を介する細胞内情報伝達機構とその病態

Research Project

Project/Area Number	03404022
Research Institution	Kobe University
Principal Investigator	高井義美神戸大学, 医学部, 教授 (60093514)
Co-Investigator(Kenkyū-buntansha)	佐々木卓也神戸大学, 医学部, 助手 (40241278) 菊池章神戸大学, 医学部, 講師 (10204827) 貝淵弘三神戸大学, 医学部, 助教授 (00169377)
Keywords	低分子量GTP結合蛋白質 / ras p21 / rho p21 / smg p25A / rab3A p25 / smg GDS / rho GDI / MAPキナーゼ / rabphilin
Research Abstract	最近の私共や他の研究室の解析により、ras p21が細胞の増殖や分化を、rho p21が細胞骨格を、smg p25A/ra3A p25が分泌反応を制御していることが明らかになりつつある。本年度、私共はこれらの低分子量GTP結合蛋白質(G蛋白質)の機能と活性化機構の解析を行い、以下の成果を得た。ras p21は細胞レベルでMAPキナーゼを活性化することが報告されていたが、私共は、ツメガエル卵の細胞質画分にKi-ras p21に依存してMAPキナーゼを活性化する因子が存在することを明らかにし、これを部分精製してREKSと命名した。rho p21に関しては、私共は、すでにrho p21に作用する活性制御蛋白質としてsmg GDSとrho GDIを見出している。本年度、私共は、rho p21の活性化を阻害するrho GDIを種々の細胞へゲイクロインジェクションすることにより、rho p21が細胞形態の維持や細胞運動、細胞質分裂を制御していることを明らかにした。また、私共は、このrho GDIがsmg GDSの作用に打ち勝ってrho p21に作用することも明らかにした。さらに、生体内でもrho p21はsmg GDSではなくrho GDIと複合体を形成していることを見出した。またrab3A p25に関しては、rab3A p25の標的蛋白質rabphilinをウシ大脳に見出し、そのcDNAをクローニングして一次構造を決定した。以上の結果を得たことにより、ras p21,rho p21およびrab3A p25の機能と活性化機構について着実な成果をあげることができ、本年度の研究計画はほぼ達成できたと考えている。

Research Products

(30 results)

All Other

All Publications (30 results)

[Publications] Miura,Y.: "Regulation of morphology by rho p21 and its inhibitory GDP/GTP exchange protein (rho GDI) in Swiss 3T3 cells." J.Biol.Chem.268. 510-515 (1993)
[Publications] Itoh,T.: "The post-translationally processeeing of ras p21 is critical for its stimulation of Mitogen-activated protein (MAP) kinase." J.Biol.Chem.(1993)
[Publications] Takaishi,K.: "Involvement of rho p21 and its inhibitory GDP/GTP exchange protein (rho GDI) in cell motility." Mol.Cell.Biol.13. 72-79 (1993)
[Publications] Shirataki,H.: "Rabphilin-3A, a putative target protein for smg p25A/rab3A p25 small GTP-binding protein related to synaptotagmin." Mol.Cell.Biol.(1993)
[Publications] Kishi,K.: "Regulation of cytoplasmic devesion of Xenopus embryo by rho p21 and its inhibitory GDP/GTP exchange protein (rho GDI)." J.Cell Biol.(1993)
[Publications] Itoh,T.: "A novel protein factor for ras p21-dependent activation of Mitogen-activated protein kinase." Proc.Natl.Acad.Sci.USA. 90. 975-979 (1993)
[Publications] Kawamura,M.: "Translocation of Ki-ras P21 between membrane and cytoplasm by smg GDS." Biochem.Biophys.Res.Commun.(1993)
[Publications] Ando,S.: "Post-translational processing of rac p21s is important both for their interaction with the GDP/GTP exchange proteins and for their activation of NADPH oxidase." J.Biol.Chem.267. 25709-25713 (1992)
[Publications] Regazzi,R.: "The small GTP-binding proteins in the cytosol of insulin-secreting cells are complexed to GDP dissociation inhibitor proteins." J.Biol.Chem.267. 17512-17519 (1992)
[Publications] Kikuchi,A.: "Functional interactions of stimulatory and inhibitory GDP/GTP exchange proteins and their common substrate small GTP-binding protein." J.Biol.Chem.267. 14611-14615 (1992)
[Publications] Shirataki,H.: "A possible target protein for smg-25A/rab3A small GTP-binding protein." J.Biol.Chem.267. 10946-10949 (1992)
[Publications] Mizuno,T.: "Regulation of the superoxide-generating NADPH oxidase by a small GTP-binding protein and its stimulatory and inhibitory GDP/GTP exchange proteins." J.Biol.Chem.267. 10215-10218 (1992)
[Publications] Musha,T.: "The geranylgeranyl moiety but not the methyl moiety of the smg-25A/rab3A protein is essential for the interactions with membrane and its inhibitory GDP/GTP exchange protein." J.Biol.Chem.267. 9821-9825 (1992)
[Publications] Hirata,K.: "Involvement of rho p21 in the GTP-enhanced calcium ion sensitivity of smooth muscle contraction." J.Biol.Chem.267. 8719-8722 (1992)
[Publications] Sugai,M.: "Epidermal cell differentiation inhibitor ADP-ribosylates small GTP-binding proteins and induces hyperplasia in epidermis." J.Biol.Chem.267. 2600-2604 (1992)
[Publications] Fujioka,H.: "Transforming and c-fos promoter/enhancerstimulating activities of a stimulatory GDP/GTP exchange protein for small GTP-binding proteins." J.Biol.Chem.267. 926-930 (1992)
[Publications] Horiuchi,H.: "The posttranslational processing of ras p21 is critical for its stimulation of yeast adenylate cyclase." Mol.Cell.Biol.12. 4515-4520 (1992)
[Publications] Yoshida,Y.: "Microinjention of smg/rap1/Krev-1 p21 into Swiss 3T3 cells induces DNA synthesis and morphological changes." Mol.Cell.Biol.12. 3407-3414 (1992)
[Publications] Ruggieri,R.: "RSR1,a ras-like gene homologous to Krev-1(smg21A/rap1A):Role in the development of cell polarity and interactions with the Ras pathway in Saccharomyces cerevisiae." Mol.Cell.Biol.12. 758-766 (1992)
[Publications] Sakoda,T.: "smg/rap1/Krev-1 p21s inhibit the signal pathway to the c-fos promoter/enhancer from c-Ki-ras p21 bus not from c-raf-1 kinase in NTH3T3 cells." Oncogene. 7. 1705-1711 (1992)
[Publications] Kotani,K.: "The functional domain of the stimulatory GDP/GTP exchange protein (smg GDS) which interacts with the C-terminal geranylgeranylated region of rap1/Krev-1/smg p21." Oncogene. 7. 1699-1704 (1992)
[Publications] Adachi,M.: "Induction of smg p21/rap1A p21/Krev-1 p21 gene expression during phorbol ester-induced differentation of a human megakaryocytic leukemia cell line." Oncogene. 7. 323-329 (1992)
[Publications] Kikuchi,A.: "Molecular cloning of the human cDNA a stimulatory GDP/GTP exchange protein for c-Ki-ras p21 and smg p21." Oncogene. 7. 289-293 (1992)
[Publications] Miura,Y.: "Phosphorylation of smg p21B/rap1B p21 by cyclic GMP-dependent protein kinase." FEBS Lett.297. 171-174 (1992)
[Publications] Mizoguchi,A.: "Localization of smg p25A/rab3A p25, a small GTP-binding protein, at the active zone of the rat neuromuscular junction." Biochem.Biophys.Res.Commun.186. 1345-1352 (1992)
[Publications] Kuroda,S.: "Cooperative function of rho GDS and rho GDI to regulate rho p21 activation in smooth muscle." Biochem.Biophys.Res.Commun.185. 473-480 (1992)
[Publications] Kim,Y.S.: "Effect of chlorpromazine on the smg GSD action." Biochem.Biophys.Res.Commun.182. 1446-1453 (1992)
[Publications] Hiraoka,K.: "Both stimulatory and inhibitory GDP/GTP exchange proteins, smg GDS and rho GDI, are active on multiple small GTP-binding proteins." Biochem.Biophys.Res.Commun.182. 921-930 (1992)
[Publications] Regazzi,R.: "Characterization of small-molecular-mass guaninenucleotide-binding regulatory proteins in insulin-secreting cells and PC12 cells." Eur.J.Biochem.208. 729-737 (1992)
[Publications] Takai,Y.: "Small GTP-binding proteins." Int.Rev.Cytol.133. 187-230 (1992)

1992 Fiscal Year Annual Research Report

低分子量GTP結合蛋白質を介する細胞内情報伝達機構とその病態

Principal Investigator

高井 義美 神戸大学, 医学部, 教授 (60093514)

Research Products

[Publications] Miura,Y.: "Regulation of morphology by rho p21 and its inhibitory GDP/GTP exchange protein (rho GDI) in Swiss 3T3 cells." J.Biol.Chem.268. 510-515 (1993)

[Publications] Itoh,T.: "The post-translationally processeeing of ras p21 is critical for its stimulation of Mitogen-activated protein (MAP) kinase." J.Biol.Chem.(1993)

[Publications] Takaishi,K.: "Involvement of rho p21 and its inhibitory GDP/GTP exchange protein (rho GDI) in cell motility." Mol.Cell.Biol.13. 72-79 (1993)

[Publications] Shirataki,H.: "Rabphilin-3A, a putative target protein for smg p25A/rab3A p25 small GTP-binding protein related to synaptotagmin." Mol.Cell.Biol.(1993)

[Publications] Kishi,K.: "Regulation of cytoplasmic devesion of Xenopus embryo by rho p21 and its inhibitory GDP/GTP exchange protein (rho GDI)." J.Cell Biol.(1993)

[Publications] Itoh,T.: "A novel protein factor for ras p21-dependent activation of Mitogen-activated protein kinase." Proc.Natl.Acad.Sci.USA. 90. 975-979 (1993)

[Publications] Kawamura,M.: "Translocation of Ki-ras P21 between membrane and cytoplasm by smg GDS." Biochem.Biophys.Res.Commun.(1993)

[Publications] Ando,S.: "Post-translational processing of rac p21s is important both for their interaction with the GDP/GTP exchange proteins and for their activation of NADPH oxidase." J.Biol.Chem.267. 25709-25713 (1992)

[Publications] Regazzi,R.: "The small GTP-binding proteins in the cytosol of insulin-secreting cells are complexed to GDP dissociation inhibitor proteins." J.Biol.Chem.267. 17512-17519 (1992)

[Publications] Kikuchi,A.: "Functional interactions of stimulatory and inhibitory GDP/GTP exchange proteins and their common substrate small GTP-binding protein." J.Biol.Chem.267. 14611-14615 (1992)

[Publications] Shirataki,H.: "A possible target protein for smg-25A/rab3A small GTP-binding protein." J.Biol.Chem.267. 10946-10949 (1992)

[Publications] Mizuno,T.: "Regulation of the superoxide-generating NADPH oxidase by a small GTP-binding protein and its stimulatory and inhibitory GDP/GTP exchange proteins." J.Biol.Chem.267. 10215-10218 (1992)

[Publications] Musha,T.: "The geranylgeranyl moiety but not the methyl moiety of the smg-25A/rab3A protein is essential for the interactions with membrane and its inhibitory GDP/GTP exchange protein." J.Biol.Chem.267. 9821-9825 (1992)

[Publications] Hirata,K.: "Involvement of rho p21 in the GTP-enhanced calcium ion sensitivity of smooth muscle contraction." J.Biol.Chem.267. 8719-8722 (1992)

[Publications] Sugai,M.: "Epidermal cell differentiation inhibitor ADP-ribosylates small GTP-binding proteins and induces hyperplasia in epidermis." J.Biol.Chem.267. 2600-2604 (1992)

[Publications] Fujioka,H.: "Transforming and c-fos promoter/enhancerstimulating activities of a stimulatory GDP/GTP exchange protein for small GTP-binding proteins." J.Biol.Chem.267. 926-930 (1992)

[Publications] Horiuchi,H.: "The posttranslational processing of ras p21 is critical for its stimulation of yeast adenylate cyclase." Mol.Cell.Biol.12. 4515-4520 (1992)

[Publications] Yoshida,Y.: "Microinjention of smg/rap1/Krev-1 p21 into Swiss 3T3 cells induces DNA synthesis and morphological changes." Mol.Cell.Biol.12. 3407-3414 (1992)

[Publications] Ruggieri,R.: "RSR1,a ras-like gene homologous to Krev-1(smg21A/rap1A):Role in the development of cell polarity and interactions with the Ras pathway in Saccharomyces cerevisiae." Mol.Cell.Biol.12. 758-766 (1992)

[Publications] Sakoda,T.: "smg/rap1/Krev-1 p21s inhibit the signal pathway to the c-fos promoter/enhancer from c-Ki-ras p21 bus not from c-raf-1 kinase in NTH3T3 cells." Oncogene. 7. 1705-1711 (1992)

[Publications] Kotani,K.: "The functional domain of the stimulatory GDP/GTP exchange protein (smg GDS) which interacts with the C-terminal geranylgeranylated region of rap1/Krev-1/smg p21." Oncogene. 7. 1699-1704 (1992)

[Publications] Adachi,M.: "Induction of smg p21/rap1A p21/Krev-1 p21 gene expression during phorbol ester-induced differentation of a human megakaryocytic leukemia cell line." Oncogene. 7. 323-329 (1992)

[Publications] Kikuchi,A.: "Molecular cloning of the human cDNA a stimulatory GDP/GTP exchange protein for c-Ki-ras p21 and smg p21." Oncogene. 7. 289-293 (1992)

[Publications] Miura,Y.: "Phosphorylation of smg p21B/rap1B p21 by cyclic GMP-dependent protein kinase." FEBS Lett.297. 171-174 (1992)

[Publications] Mizoguchi,A.: "Localization of smg p25A/rab3A p25, a small GTP-binding protein, at the active zone of the rat neuromuscular junction." Biochem.Biophys.Res.Commun.186. 1345-1352 (1992)

[Publications] Kuroda,S.: "Cooperative function of rho GDS and rho GDI to regulate rho p21 activation in smooth muscle." Biochem.Biophys.Res.Commun.185. 473-480 (1992)

[Publications] Kim,Y.S.: "Effect of chlorpromazine on the smg GSD action." Biochem.Biophys.Res.Commun.182. 1446-1453 (1992)

[Publications] Hiraoka,K.: "Both stimulatory and inhibitory GDP/GTP exchange proteins, smg GDS and rho GDI, are active on multiple small GTP-binding proteins." Biochem.Biophys.Res.Commun.182. 921-930 (1992)

[Publications] Regazzi,R.: "Characterization of small-molecular-mass guaninenucleotide-binding regulatory proteins in insulin-secreting cells and PC12 cells." Eur.J.Biochem.208. 729-737 (1992)

[Publications] Takai,Y.: "Small GTP-binding proteins." Int.Rev.Cytol.133. 187-230 (1992)

高井義美神戸大学, 医学部, 教授 (60093514)