• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to project page

2000 Fiscal Year Final Research Report Summary

Mechanism of Intracellular Signaling via Ras Protein

Research Project

Project/Area Number 06280218
Research Category

Grant-in-Aid for Scientific Research on Priority Areas

Allocation TypeSingle-year Grants
Research InstitutionKobe University School of Medicine

Principal Investigator

KATAOKA Tohru  Kobe Univ.Sch.Med., Dept.Physiology II, Professor, 医学部, 教授 (40144472)

Co-Investigator(Kenkyū-buntansha) OKADA Tomoyo  Kobe Univ.Sch.Med., Dept.Physiology II, Instructor, 医学部, 助手 (80294205)
HU Chang-deng  Kobe Univ.Sch.Med., Dept.Physiology II, Instructor, 医学部, 助手 (70294204)
KATAOKA Yuriko  Kobe Univ.Sch.Med., Dept.Physiology II, Instructor (50233739)
KARIYA Ken-ichi  Kobe Univ.Sch.Med., Dept.Physiology II, Associate Prof. (40263371)
Project Period (FY) 1994 – 1999
Keywordsras Oncogene / Posttranslational Modification / Farnesylation / Phospholipase C / Adenylyl Cyclase / Adenylyl Cyclase-Associated Protein / raf Oncogene / rap1 Anti-Oncogene
Research Abstract

1. We discovered a novel interaction between Ras and its effectors ; human Raf and yeast adenylyl cyclase. This "second" interaction is essential for full activation of the effectors by Ras, and dependent on the "activator" region and the posttranslational modification (famesylation) of Ras. The second Ras-binding sites are located in the cysteine-rich domain of Raf and in a complex between cyclase-associated protein CAP and the adenylyl cyclase C-terminus. Based on these findings, we elucitated the molecular mechanisms by which the anti-oncogene product Rap1 antagonizes the Ras function, by which Ras and Rap1 exert differential regulatory activities toward Raf-1 and B-Raf, and by which phosphorylation of Rap1 by A-kinase results in inhibition of its Ras-antagonizing activity. We proposed that the strength of the second interaction is a critical determinant of effector regulation by Ras family small G-proteins.
2. By screening many single amino acid substitution mutants of H-Ras for interaction with various Ras effector proteins, we demonstrated that significant differences exist in the recognition mechanisms by which the multiple effector proteins associate with Ras, and obtained H-Ras mutants that could discriminate the effectors. Using these mutants, we examined which effector was really involved in some Ras-induced cellular events.
3. We discovered a novel Ras-effector candidate PLC-ε in nematode and isolated its human homologue. The human PLC-ε exhibited GTP-dependent binding to Ras/Rap1, and, when coexpressed with Ras and Rap1, was translocated from the cytoplasm to the plasma membrane and the Golgi apparatus, respectively, and had its phospholipase C activity greatly activated. Thus, PLC-ε is a real Ras effector. In addition, we discovered two novel proteins RA-GEF-1 and-2, which possessed both a RA domain associating directly with the GTP-bound Rap1 and a GEF domain catalysing GDP-GTP exchange on Rap1.

  • Research Products

    (47 results)

All Other

All Publications (47 results)

  • [Publications] Minato,T., et al.: "Quantitative analysis of the mutually competitive binding of human Raf-1 and yeast adenylyl cyclase to Ras proteins"J.Biol.Chem.. 269. 20845-20851 (1994)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Masuda,T., et al.: "Protein kinase Byr2 is a target of Ras1 in fission yeast Schizosaccharomyces pombe"J.Biol.Chem.. 270. 1979-1982 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hu,C.-D., et al.: "Cysteine-rich region or Raf-1 interacts with activator domain of post-translationally modified H-Ras"J.Biol.Chem.. 270. 30274-30277 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Okada,T., et al.: "Post-translational modification of H-Ras is required for activation of, but not for association with, B-Raf"J.Biol.Chem.. 271. 4671-4678 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Akasaka,K., et al.: "Differential Structural requirements for interaction of Ras protein with its distinct downstream effectors"J.Biol.Chem.. 271. 5353-5360 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Shinkai,M., et al.: "Difference i n the mechanism of interaction of Raf-1 and B-Raf with H-Ras"Biochem.Biophys.Res.Comm.. 223. 729-734 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Shima,F., et al.: "Effect of association with adenylyl cyclase-associated protein on the interaction of yeast adenylyl cyclase with Ras protein"Mol.Cell.Biol.. 17. 1057-1064 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Okazaki,M., et al.: "Synergistic activation of c-fos proinoter activity by Raf and Ral GDP dissociation stimulator"Oncogene. 14. 515-521 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yanagihara,C., et al.: "Association of elongation factor 1α and ribosomal protein L3 with the proline-rich region of yeast adenylyl cyclase-associated protein CAP"Biochem.Biophys.Res.Comm.. 232. 503-507 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hu,C-D., et al.: "Coassociation of Rap1A and Ha-Ras with Raf-1 N-terminal region interferes with Ras-dependent activation of Raf-1"J.Biol.Chem.. 272. 11702-11705 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Tamada,M., et al.: "Membrane recruitment of Raf-1 is not the only function of Ras i n Raf-1 activation"Oncogene. 15. 2959-2964 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Shibatohge,M., et al.: "Identification of PLC210, a Caenorhabditis elegans phospholipase C, as a putative effector of Ras"J.Biol.Chem.. 273. 6218-6222 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Shirouzu,M., et al.: "Interactions of the amino acid residue at position 31 of the c-Ha-Ras protein with Raf-1 and RalGDS"J.Biol.Chem.. 273. 7737-7742 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ohnishi,M., et al.: "Selective inhibition of Ras interaction with its particular effector by synthetic peptides corresponding to the Ras effector region"J.Biol.Chem.. 273. 10210-10215 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kim,J.H., et al.: "Activation of Ras and its downstream extracellular signal-regulated protein kinases by the CDC25 homology domain of mouse Son-of-sevenless 1 (mSos1)"Oncogene. 16. 2597-2607 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Nishida,Y., et al.: "Coiled-coil interaction of N-terminal 36 residues of cyclase-associated protein with adenylyl cyclase is sufficient for its function in Saccharomyces cerevisiae Ras pathway"J.Biol.Chem.. 273. 28019-28024 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Watari,Y., et al.: "Identification of Ce-AF-6, a novel Caenorhabditis elegans protein, as a Ras putative effector"Gene. 224. 53-58 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hu,C-D., et al.: "Effect of phosphorylation on activities of Rap1A to interact with Raf-1 and to suppress Ras-dependent Raf-1 activation"J.Biol.Chem.. 274. 48-51 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Goshima,M., et al.: "Characterization of a novel Ras-binding protein Ce-FLI-1 comprising leucine-rich repeats and gelsolin-like domains"Biochem.Biophys.Res.Comm.. 257. 111-116 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Okada,T., et al.: "The strength of interaction at the Raf cysteine-rich domain is a critical determinant of response of Raf to Ras family small GTPases"Mol.Cell.Biol.. 19. 6057-6064 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Tanaka,Y., et al.: "H-Ras signals to cytoskeletal machinery in induction of integrin-mediated adhesion of T cells"J.Immunol.. 163. 6209-6216 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Liao,Y., et al.: "RA-GEF, a novel Rap1A guanine nucleotide exchange factor containing a Ras/Rap1A-associating domain, is conserved between nematode and humans"J.Biol.Chem.. 274. 37815-37820 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Shima,F., et al.: "Association of yeast adenylyl cyclase with cyclase-associated protein CAP forms a second Ras-binding site which mediates its Ras-dependent activation"Mol.Cell.Biol.. 20. 26-33 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kataoka,T., et al.: "Cytoskeleton and G-proteins in the regulation of cancer"Noboru Kuzumaki. 184 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T.Minato et al.: "Quantitative analysis of the mutually competitive binding of human Raf-1 and yeast adenylyl cyclase to Ras proteins"J.Biol.Chem.. 269. 20845-20851 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T.Masuda et al.: "Protein kinase Byr2 is a target of Ras1 in fission yeast Schizosaccharomyces pombe"J.Biol.Chem.. 270. 1979-1982 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] C-D.Hu et al.: "Cysteine-rich region of Raf-1 interacts with activator domain of post-translationally modified H-Ras"J.Biol.Chem.. 270. 30274-30277 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T.Okada et al.: "Post-translational modification of H-Ras is required for activation of, but not for association with, B-Raf"J.Biol.Chem.. 271. 4671-4678 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] K.Akasaka et al.: "Differential structural requirements for interaction of Ras protein with its distinct downstream effectors"J.Biol.Chem.. 271. 5353-5360 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] M.Shinkai et al.: "Difference in the mechanism of interaction of Raf-1 and B-Raf with H-Ras"Biochem.Biophys.Res.Comm. 223. 729-734 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] F.Shima et al.: "Effect of association with adenylyl cyclase-associated protein on the interaction of yeast adenylyl cyclase with Ras protein"Mol.Cell.Biol.. 17. 1057-1064 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] M.Okazaki et al.: "Synergistic activation of c-fos promoter activity by Raf and Ral GDP dissociation stimulator"Oncogene. 14. 515-521 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] C.Yanagihara et al.: "Association of elongation factor 1α and ribosomal protein L3 with the proline-rich region of yeast adenylyl cyclase-associated protein CAP"Biochem.Biophys.Res.Comm.. 232. 503-507 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] C-D.Hu et al.: "Coassociation of Rap1 A and Ha-Ras with Raf-1 N-terminal region interferes with Ras-dependent activation of Raf-1"J.Biol.Chem.. 272. 11702-11705 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] M.Tamada et al.: "Membrane recruitment of Raf-1 is not the only function of Ras in Raf-1 activation."Oncogene. 15. 2959-2964 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] M.Shibatohge et al.: "Identification of PLC210, a Caenorhabditis elegans phospholipase C, as a putative effector of Ras."J.Biol.Chem.. 273. 6218-6222 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] M.Shirouzu et al.: "Interactions of the amino acid residue at position 31 of the c-Ha-Ras protein with Raf-1 and RalGDS."J.Biol.Chem.. 273. 7737-7742 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] M.Ohnishi et al.: "Selective inhibition of Ras interaction with its particular effector by synthetic peptides corresponding to the Ras effector region."J.Biol.Chem.. 273. 10210-10215 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] J.H.Kim et al.: "Activation of Ras and its downstream extracellular signal-regulated protein kinases by the CDC25 homology domain of mouse Son-of-sevenless 1 (mSos 1)"Oncogene. 16. 2597-2607 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Y.Nishida et al.: "Coiled-coil interaction of N-terminal 36 residues of cyclase-associated protein with adenylyl cyclase is sufficient for its function in Saccharomyces cerevisiae Ras pathway."J.Biol.Chem.. 273. 28019-28024 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Y.Watari et al.: "Identification of Ce-AF-6, a novel Caenorhabditis elegans protein, as a Ras putative effector"Gene. 224. 53-58 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] C-D.Hu et al.: "Effect of phosphorylation on activities of Rap 1 A to interact with Raf-1 and to suppress Ras-dependent Raf-1."J.Biol.Chem.. 274. 48-51 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] M.Goshima et al.: "Characterization of a novel Ras-binding protein Ce-FLI-l comprising leucine-rich repeats and gelsolin-like domains"Biochem.Biophys.Res.Comm.. 257. 111-116 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T.Okada et al.: "The strength of interaction at the Raf cysteine-rich domain is a critical determinant of response of Raf to Ras family small GTPases"Mol.Cell.Biol.. 19. 6057-6064 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Y.Tanaka et al.: "H-Ras signals to cytoskeletal machinery in induction of integrin-mediated adhesion of T cells"J.Immunol.. 163. 6209-6216 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Y.Liao et al.: "RA-GEF, a novel Rap 1 A guanine nucleotide exchange factor containing a Ras/Rap 1 A-associating domain, is conserved between nematode and humans"J.Biol.Chem.. 274. 37815-37820 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] F.Shima et al.: "Association of yeast adenylyl cyclase with cyclase-associated protein CAP forms a second Ras-binding site which mediates its Ras-dependent activation"Mol.Cell.Biol.. 20. 26-33 (2000)

    • Description
      「研究成果報告書概要(欧文)」より

URL: 

Published: 2002-03-26  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi