細胞内情報伝達系と細胞増殖の制御機構

• 研究課題/領域番号: 04253229
• 研究種目: 重点領域研究
• 配分区分: 補助金
• 研究機関: 神戸大学
• 研究代表者: 高井 義美 神戸大学, 医学部, 教授 (60093514)
• 研究分担者: 田中 一馬 神戸大学, 医学部, 助手 (60188290) ; 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)
• 研究期間 (年度): 1993
• 研究課題ステータス: 完了 (1993年度)
• 配分額: 52,000千円 (直接経費: 52,000千円); 1993年度: 27,000千円 (直接経費: 27,000千円); 1992年度: 25,000千円 (直接経費: 25,000千円)
• キーワード: 低分子量G蛋白質 / Rho / HGF / 細胞運動 / Rho GDI / C3 / TPA感受性因子 / Cキナーゼ / 細胞増殖 / 低分子量GTP結合蛋白質 / ras p21 / rho p21 / smg GDS / rho GDI

研究概要

低分子量G蛋白質の中でも、Rasは核遺伝子の発現を介して細胞の増殖や分化を制御していることが判明しているが、私共は本研究でRho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を明らかにした。すなわち、ケラチノサイト308R細胞にRhoの活性制御蛋白質であるRho GDIや、RhoをADPリボシル化してその機能を阻害するC3酵素をマイクロインジェクションすると、HGFによる細胞運動が完全に阻害された。このRho GDIやC3の作用は、活性型のRhoやADPリボシル化をうけない変異型Rhoと共にインジェクションすると認められなかった。また、Cキナーゼを活性化するTPAを作用させても細胞運動が引き起こされ、この作用はRho GDIやC3によって阻害された。細胞運動はRasやCa^<2+>とは直接関係していなかった。これらの結果から、HGFの受容体Metは何らかの機構でTPA感受性因子(Cキナーゼ?)を介してRhoを活性化し、その結果、細胞運動が引き起こされると考えられる。現在、TPA感受性因子がどのような機構でRhoの活性を制御しているか、Rhoがどのような機構で細胞運動を制御しているかを解析中である。Rasやその関連蛋白質の遺伝子の異常が細胞の異常増殖を引き起こすことはすでに判明しているが、これらのアナロジーからRhoとその関連蛋白質の遺伝子の異常ががん細胞の浸潤・転移に関与している可能性がでてきた。現在、RhoやRho関連蛋白質の遺伝子が浸潤・転移度の高いがん細胞で異常があるか否かも検討している。一方、Rasの活性制御蛋白質と標的蛋白質であるSmg GDS,REKSは共に私共の研究室で発見したものであるが、これらに関する本年度の研究も順調に進んだ。

研究成果

• [文献書誌] 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)
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• [文献書誌] Kishi,K.: "Regulation of cytoplasmic division of Xenopus embryo by rho p21 and its inhibitory GDP/GTP exchange protein(rho GDI)." J.Cell Biol.120. 1187-1195 (1993)
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• [文献書誌] Yaku,H.: "The Dbl oncogene product as a GDP/GTP exchange protein for the Rho family.Its properties in comparison with those of Smg GDS" Biochem.Biophys.Res.Commun.198. 811-818 (1994)
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• [文献書誌] 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)
• [文献書誌] 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)
• [文献書誌] Shirataki,H.: "Rabphilin-3A,aputative target protein for smgp25A/rab3A p25 small GTP-binding protein related to synaptotagmin." Mol.Cell.Biol.in press. (1993)
• [文献書誌] 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)
• [文献書誌] 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)
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• [文献書誌] Horiuchi,H.: "The posttranslational processing of ras p21 is critical for its stimulation of yeast adenylate cyclade." Mol.Cell.Biol. 12. 4515-4520 (1992)
• [文献書誌] 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)
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• [文献書誌] 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)
• [文献書誌] 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)
• [文献書誌] 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)
• [文献書誌] 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)
• [文献書誌] Miura,Y.: "Phosphorylation of smg p21B/rap1B p21 by cyclic GMP-dependent protein kinade." FEBA Lett.297. 171-174 (1992)
• [文献書誌] 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)
• [文献書誌] 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)
• [文献書誌] Kim,Y.S.: "Effect of chlorpromazine on the smg GDS acton." Biochem.Biophys.Res.Commun.182. 1446-1453 (1992)
• [文献書誌] 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)
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