1998 Fiscal Year Annual Research Report

低分子量G蛋白質による細胞の増殖、運動の制御機構

Research Project

Project/Area Number	09254103
Research Institution	Hiroshima University
Principal Investigator	菊池章広島大学, 医学部, 教授 (10204827)
Co-Investigator(Kenkyū-buntansha)	大矢禎一東京大学大学院, 理学系研究科, 助教授 (20183767) 松井泰東京大学大学院, 理学系研究科, 助手 (50229407) 松田道行国立国際医療センター研究所, 臨床病理研究部, 部長 (10199812) 服部成介国立精神神経センター神経研究所, 診断研究部, 室長 (50143508) 佐藤孝哉東京工業大学, 生命理工学部, 寄附講座教員 (20251655)
Keywords	Ras / Ral / RasGRF1 / GAP1m / CalDAGI / cAMP-GEF / Rho / PLD
Research Abstract	低分子量G蛋白質の中でも発がん、転移に重要であるRasとRhoの活性化機構と作用機構について解析を行った。Ras-RalGDS-Ralの下流で機能する新規蛋白質POB1とEpsinを見出した。RalやRalBP1(Ralの標的蛋白質)、POB1(RalBP1の結合蛋白質)、Epsin(POB1の結合蛋白質)は、いずれもインスリンとEGF依存性のエンドサイトーシスを制御した。Ras-GRF1が三量体G蛋白質βγサブユニットの刺激により、RacのGDP/GTP交換促進因子として機能し、この活性にはDb1ドメインが必要であった。三量体G蛋白質G12のαサブユニットがGTP依存性にGap1mのPHドメインに結合し、Gap1mのGAP活性を促進し、その結果、EGFによるRasの活性化を抑制した。Rap1の新規活性化囚子CalDAGIとcAMP-GEFを同定した。CalDAGIのRap1に対する活性は、Ca^<2+>およびDACにより活性化された。また、cAMP-GEFはサイクリックAMPにより活性化されされた。酵母Rho3は、極性分泌を制御するExocyst複合体のコンポーネントであるExo70とミオシンVであるMyo2とに結合し、Exo70の細胞内局在がRho3により変化した。酵母Rho1pの標的酵素である1,3-β-グルカン合成酵素の温度感受性変異株のうちで、推定上の基質結合部位近傍に変異を持つ4つの変異株では、in vitroでのグルカン合成酵素活性が特に制限温度下で顕著に低下していて、多くの細胞が小さな芽様の突起を出して増殖を停止した。PLD1とPLD2はそれぞれ後期エンドソーム/リソゾームと細胞膜/オルガネラ膜系に局在し、EGF刺激により、PLD1の局在は変化しないが、PLD2はラッフリング膜に濃縮された。

Research Products

(18 results)

All Other

All Publications (18 results)

[Publications] Ikeda,M.: "Identification and characterization of a novel protein interacting with Ral-binding proteinl,a putative effector protein of Ral." J.Biol.Chem.273・2. 814-821 (1998)
[Publications] Shirouzu,M.: "Interactions of the amino acid residues at position 31 of the c-Ha-Ras protein with Raf-1 and RalGDS." J.Biol.Chem.273・13. 7737-7742 (1998)
[Publications] Deng,J.: "Involvement of Ras in Bruton's tyrosine kinase-mediated JNK activation." J.Biol.Chem.273・27. 16787-16791 (1998)
[Publications] Jiang,Y.: "The G protein G12 stimulates Bruton's tyrosine kinase and a rasGAP through a conserved PH/BM domain" Nature. 395・6704. 808-813 (1998)
[Publications] Matsumura,I.: "Involvement of prolonged ras activation in thrombopoietin-induced megakaryocytic differentiantion of a human factor-dependent hematopoietic cell line." Mol.Cell.Biol. 18・7. 4282-4290 (1998)
[Publications] Hashimoto,Y.: "Phosphorylation of CrkII adaptor protein at Tyrosine 221 by epidermal growth factor receptor." J.Biol.Chem.273・27. 17186-17191 (1998)
[Publications] Okada,S.: "Insulin regulates the dynamic balance batween Ras and Rapl signaling by coordinating the assembly states of the Grb2-SOS and CrkII-C3G complexes." EMBO J.17・9. 2554-2565 (1998)
[Publications] Kawasaki,H.: "A family of cAMP-binding proteins that directly activate Rapl." Science. 282・5397. 2275-2279 (1998)
[Publications] Caplin B.E.: "Amino acid residues that define both the isoprenoid and CAAX preferences of the Saccharomyces cerevisiae protein farnesyltransferase.creating the perfect farnesyltransferase" J.Biol.Chem.273・16. 9472-9479 (1998)
[Publications] Homma,K.: "Phosphatidylinositol-4-phosphate 5-kinase localized on the plasma membrane is essential for yeast cell morphogenesis." J.Biol.Chem.273・25. 15779-15786 (1998)
[Publications] Okano,H.: "Importance of phenylalanine residues of yeast calmodulin for target binding and activation." J.Biol.Chem.273・41. 26375-26382 (1998)
[Publications] Kagami,M.: "Sro7p,a Saccharomyces cerevisiae counterpart of the tumor suppressor l (2) g1,is functionally realted to myosines." Genetics. 149・4. 1717-1727 (1998)
[Publications] Yokozeki,T.: "Purification and characterization of phosphatidic acid-regulated protein kinase and its substrate,p29." J.Neurochem.71・1. 410-417 (1998)
[Publications] Matsubara,K.: "Plasma membrane recruitment of RalGDS is critical for Ras-dependent Ral Activation." Oncogene. 18・6. 1303-1312 (1999)
[Publications] Nishida,K.: "Anti-apoptotic function of Rac in hematopoietic cells." Oncogene. 18・2. 407-415 (1999)
[Publications] Sawamoto,K.: "Ectopic expression of mutationally-activated RalGTPase inhibits cellshape changes during Drosophila eye development." Oncogene. (in press). (1999)
[Publications] Robinson,N.: "Rho3 of Saccharomyces cerevisiae,which regulates the actin cytoskeleton and exocytosis,is a GTPase which interacts with Myo2 and Exo70." Mol.Cell.Biol.(in press). (1999)
[Publications] Dohke,Y.: "Translocation of Arfl to the secretory granules in rat parotid acinar cells." Arch.Biochem.Biophys.(in press). (1999)

1998 Fiscal Year Annual Research Report

低分子量G蛋白質による細胞の増殖、運動の制御機構

Principal Investigator

菊池 章 広島大学, 医学部, 教授 (10204827)

Research Products

[Publications] Ikeda,M.: "Identification and characterization of a novel protein interacting with Ral-binding proteinl,a putative effector protein of Ral." J.Biol.Chem.273・2. 814-821 (1998)

[Publications] Shirouzu,M.: "Interactions of the amino acid residues at position 31 of the c-Ha-Ras protein with Raf-1 and RalGDS." J.Biol.Chem.273・13. 7737-7742 (1998)

[Publications] Deng,J.: "Involvement of Ras in Bruton's tyrosine kinase-mediated JNK activation." J.Biol.Chem.273・27. 16787-16791 (1998)

[Publications] Jiang,Y.: "The G protein G12 stimulates Bruton's tyrosine kinase and a rasGAP through a conserved PH/BM domain" Nature. 395・6704. 808-813 (1998)

[Publications] Matsumura,I.: "Involvement of prolonged ras activation in thrombopoietin-induced megakaryocytic differentiantion of a human factor-dependent hematopoietic cell line." Mol.Cell.Biol. 18・7. 4282-4290 (1998)

[Publications] Hashimoto,Y.: "Phosphorylation of CrkII adaptor protein at Tyrosine 221 by epidermal growth factor receptor." J.Biol.Chem.273・27. 17186-17191 (1998)

[Publications] Okada,S.: "Insulin regulates the dynamic balance batween Ras and Rapl signaling by coordinating the assembly states of the Grb2-SOS and CrkII-C3G complexes." EMBO J.17・9. 2554-2565 (1998)

[Publications] Kawasaki,H.: "A family of cAMP-binding proteins that directly activate Rapl." Science. 282・5397. 2275-2279 (1998)

[Publications] Caplin B.E.: "Amino acid residues that define both the isoprenoid and CAAX preferences of the Saccharomyces cerevisiae protein farnesyltransferase.creating the perfect farnesyltransferase" J.Biol.Chem.273・16. 9472-9479 (1998)

[Publications] Homma,K.: "Phosphatidylinositol-4-phosphate 5-kinase localized on the plasma membrane is essential for yeast cell morphogenesis." J.Biol.Chem.273・25. 15779-15786 (1998)

[Publications] Okano,H.: "Importance of phenylalanine residues of yeast calmodulin for target binding and activation." J.Biol.Chem.273・41. 26375-26382 (1998)

[Publications] Kagami,M.: "Sro7p,a Saccharomyces cerevisiae counterpart of the tumor suppressor l (2) g1,is functionally realted to myosines." Genetics. 149・4. 1717-1727 (1998)

[Publications] Yokozeki,T.: "Purification and characterization of phosphatidic acid-regulated protein kinase and its substrate,p29." J.Neurochem.71・1. 410-417 (1998)

[Publications] Matsubara,K.: "Plasma membrane recruitment of RalGDS is critical for Ras-dependent Ral Activation." Oncogene. 18・6. 1303-1312 (1999)

[Publications] Nishida,K.: "Anti-apoptotic function of Rac in hematopoietic cells." Oncogene. 18・2. 407-415 (1999)

[Publications] Sawamoto,K.: "Ectopic expression of mutationally-activated RalGTPase inhibits cellshape changes during Drosophila eye development." Oncogene. (in press). (1999)

[Publications] Robinson,N.: "Rho3 of Saccharomyces cerevisiae,which regulates the actin cytoskeleton and exocytosis,is a GTPase which interacts with Myo2 and Exo70." Mol.Cell.Biol.(in press). (1999)

[Publications] Dohke,Y.: "Translocation of Arfl to the secretory granules in rat parotid acinar cells." Arch.Biochem.Biophys.(in press). (1999)

菊池章広島大学, 医学部, 教授 (10204827)