細胞内情報伝達系と細胞増殖の制御機構

1992 Fiscal Year Annual Research Report

• Project/Area Number: 04253229
• 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: 細胞増殖 / 低分子量GTP結合蛋白質 / ras p21 / rho p21 / smg GDS / rho GDI

Research Abstract

最近の私共や他の研究室の解析により、rasp21が遺伝子発現系を介して細胞の増殖や分化を、rho p21がアクトミオシン系を介して細胞の形態や運動を制御していることが明らかになりつつある。本年度は、ras p21の標的蛋白質の同定およびro p21の機能と活性化機構について解析を行い、以下の結果を得た。最近、ras p21が細胞レベルでMAPキナーゼを活性化することが報告されているが、本年度、私共は、ツメガエル卵の細胞質画分にKi-ras p21に存在して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REKSと命名して部分精製した。REKSは分子量150KDaの蛋白質で、細胞質に存在していた。低分子量G蛋白質はそのC末端に脂質による翻訳後修飾を受けることが知られているが、Ki-ras p21がREKSを活性化するのにこの翻訳後修飾が必須であった。一方、rha p21に関しては、私共は、rho p21の活性化を阻害するrha GDIを種々の細胞ヘマイクロインジュクションすることにより、rho p21が細胞運動、細胞形態や細胞質分裂を制御していることを明らかにした。私共はrho p21に作用する活性制御蛋白質としてsmg GDSとGDIを見出していたが、本年度、rho p21はこのrho GDIと細胞内で1:1の複合体を形成しており、さらに、smg GDSはこの複合体に作用できないことを明らかにした。本年度の私共の研究結果から、ras p21およびrho p21の活性制御機構とその機能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することがデきたと考えている。

Research Products

• [Publications] Miura,Y.: "Regulation of morphology by rho p21 and its inhibitory GDP/GTP exchange prorein(rho GFI)in swiss 3T3 cells." J.Biol.Chem. 268. 510-515 (1993)
• [Publications] Itoh,T.: "The post-translatioally processing of ras p21 is critical for its stimulation of Mitogen-activated protein(MAP)kinase." J.Biol.Chem.in press. (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,aputative target protein for smgp25A/rab3A p25 small GTP-binding protein related to synaptotagmin." Mol.Cell.Biol.in press. (1993)
• [Publications] Kishi,K.: "REgulation of cytoplasmic division of xemopus pmbryo by rho p21 and irs inhibitory GDP/GTP exchamge protein(rho GDI)." J.Cell Biol.in press. (1993)
• [Publications] Itoh,T.: "A novel protein factor for ras p21-dependent activation of Mitogen-activated protein kinade." Proc.Natl.Acad.Sci.USA. 90. 975-979 (1993)
• [Publications] KAwamura,M.: "Translocation of Ki-ras p21 cetween membrane and cytoplasm by smg GDS." Biochem.Biophys.Res.Commun.in press. (1993)
• [Publications] Ando,S.: "Posr-trsnslational processing of rac p21s is important both for their interaceion with the GDP/GTP exxhange proteins and for their activation of NADPH oxidade." 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 ibhibitory GDP/GTP exchamge proteins and their common substrate small GTP0-binding protein." J.Biol.Chem. 267. 14611-14615 (1992)
• [Publications] Shirataki,H.: "A possible target protein for smg-25A/rab3A small HTP-binding protein." J.Biol.Chem.267. 10946-10949 (1992)
• [Publications] Mizuno,T.: "Regulation of the superoxese-gemerating NADPH oxidase by a small BTP-bindiog and its stomulatory and inhibitory GDP/GTP exchange proteins." J.Biol.Chem.267. 10215-10218 (1992)
• [Publications] Musha,T.: "The gerany lgeranyl moiety but not the methyl molety of the smg-25A/rab3A protein is essemtial for the interactions with membrame and its inhibitory GDP/GTP exchamge protein." J.Biol.Chem.267. 9821-9825 (1992)
• [Publications] Hirata,K.: "Involvement of rho p21 in the GTP-engabced calcium ion sensitivity of smooih muscle comtraction." J.Biol.Chem.267. 8719-8722 (1992)
• [Publications] Sugai,M.: "Epidermal cell differention inhibitor ADP-ribosylates small GTP-binding proteins and induces hyperplasia in epidermis." J.Biol.Chem.267. 2600-2604 (1992)
• [Publications] Fuhioka,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 cyclade." Mol.Cell.Biol. 12. 4515-4520 (1992)
• [Publications] Yoshida,Y.: "Microinjection of smg/rap1/Krev-1 p21 into swiss 3T3 cells induces DNA synthesis and morphological chamges." 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 pathwaty in Saccharomyces cerevisiae." Mol.Cell.Biol.12. 758-766 (1992)
• [Publications] Sakod,T.: "smg/rap1/Krev-1p21s dnhibit the signal pathway to the c-fos promoter/enhancer from c-ki-ras p21but not from c-raf-1 kinase in NIH3T3 cells." Oncogene. 7. 1705-1711 (1992)
• [Publications] Kotane,K.: "Tte functional domain of the stimulatory GDP/GTP exchange protein(smg GDS)which interacts with the C-terminal geranylgerany lgerany lated region of rap1/Jrev-1/smg p21." Oncogene. 7. 1699-1704 (1992)
• [Publications] Abachi,M.: "Induction of smg p21/rap1A p21/krec-1p21 gene expression duriog phorbol ester-induced differentiation of human negakaryocytic leukemia cell line." Oncogene. 7. 323-329 (1992)
• [Publications] Kikuchi,A.: "Molecular cloning of the human cDNA for 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 kinade." FEBA 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 actuvation in smooth muscle." Biochem.Biophys.Res.Commun. 185. 473-480 (1992)
• [Publications] Kim,Y.S.: "Effect of chlorpromazine on the smg GDS acton." Biochem.Biophys.Res.Commun.182. 1446-1453 (1992)
• [Publications] Hiraoka,K.: "Both sumulatory and inhibitory GDP/GTP exxhange proteins,smg GDS and rho GDI,are active on multiple small GTP-cinding proteins." Biochem.Biophys.Res.Commun.182. 921-930 (1992)
• [Publications] Regazzi,R.: "Characterezation of small-molecular-mass guaninenucleotide-binding regulatory proteins in insulin-secretiog cells and PC12 cells." Eur.J.Biochem.208. 729-737 (1992)
• [Publications] Takai,Y.: "Small GTP-cinding proteins." Int.Rev.Cytol.133. 187-230 (1992)