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Identification and functional analysis of proteins involved in novel pathway for PLC activation.

Research Project

Project/Area Number 08458184
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Structural biochemistry
Research InstitutionFukushima Medical University

Principal Investigator

HOMMA Yoshimi  Fukushima Medical University School of Medicine, Department of Biomolecular Science, Professor, 医学部, 教授 (60192324)

Project Period (FY) 1996 – 1997
Project Status Completed (Fiscal Year 1997)
Budget Amount *help
¥6,500,000 (Direct Cost: ¥6,500,000)
Fiscal Year 1997: ¥2,600,000 (Direct Cost: ¥2,600,000)
Fiscal Year 1996: ¥3,900,000 (Direct Cost: ¥3,900,000)
Keywordsintracellular signaling / inositol phospholipid / G protein / phospholipase C / actin / cytoskeleton / イノシトールリン脂質代謝 / 細胞内セカンドメッセンジャー
Research Abstract

We recently cloned a novel signaling molecule, p122, that shows GAP activity specific for Rho and the ability to enhance the PIP_<> hydrolyzing activity of PLCdelta1 in vitro. Here we identified molecules interacting with p122 and analyzed the in vivo function of p122. Following results were obtained from this research project. 1) p122 interacted with a number of intracellular signaling molecules such as vinculin and p55.2) Microinjection of the GAP domain of p122 suppressed the formation of stress fibers and focal adhesions induced by lysophosphatidic acid. 3) Transfection of p122 also induced the morphological rounding of various adherent cells, followed by detachment from the substrate within 24 h. No stress fibers or focal adhesions were observed in the course of these changes. 4) The morphological changes caused by p122 were inhibited by coexpression of V14-RhoA, which is defective in an intrinsic GTPase activity. Analyses using deletion and point mutants demonstrated that the GAP domain of p122 is responsible for the morphological changes and detachment, and that arginine residues at positions 668 and 710 and a lysine residue at position 706 in the GAP domain of p122, all conserved among several Rho GAPs, are essential for stimulating the GTPase activity of Rho. 5) Using the Ca^<2+>-sensitive dye fluo-3, we found that microinjection of p122 evoked a rapid elevation of intracellular Ca^<2+> levels, suggesting that p122 stimulates the PIP_2 hydrolyzing activity of PLCdelta1 in vivo.

Report

(3 results)
  • 1997 Annual Research Report   Final Research Report Summary
  • 1996 Annual Research Report
  • Research Products

    (29 results)

All Other

All Publications (29 results)

  • [Publications] Homma, M. K., et al.: "Growth inhibition by phospholipase C inhibitor peptides of colorectal carcinoma oells derived from familial adenomatous polyposis." Cell Growth Differ.7. 281-288 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kabuyama, Y., et al.: "Purification and characterization of the phosphatidylinositol-3, 4, 5-trisphoshate in bovine thymus." Eur. J. Biochem.238. 350-356 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Shibata, H., et al.: "Characterization of the interaction between RhoA and the amino-terminal region of PKN." FEBS Lett.385. 214-220 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kabuyama, Y., et al.: "Wavelength specific activation of PI 3-kinase by UVB irradiation." FEBS Lett.441. 297-301 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Sekimata, M., et al.: "Morphological changes and detachment of adherent cells induced by p122." J. Biol. Chem.(印刷中).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Homma, Y & Emori, Y.: "Signalling by Inositides." Shears, S. (IRL Press Oxford), 225 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Homma Yoshimi: "Phospholipase C" Experimental Medicine (Japanese). 14. 80-82 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Homma Miwako K., Homma Yoshimi, Yamasaki Motoo, Imajoh-Ohmi Shinobu, Yuasa Yasuhito: "Growth inhibition by phospholipase C peptides of colorectal carcinoma cells derived from familial adenomatous polyposis." Cell Growth Differ.7. 281-288 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Qin Suofu, Inazu Tetsuya, Takata Minoru, Kurosaki Tomohiro, Homma Yoshimi, Yamamura Hirohei: "Cooperation of tyrosine kinase p72^<syk> and p53/56^<lyn> regulates calcium mobilization in chicken B cell oxidant stress signaling." Eur.J.Biochem.236. 443-449 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kabuyama Yukihito, Nakatsu Noriyuki, Homma Yoshimi, Fukui Yasuhisa: "Purification and characterization of the phosphatidylinositol-3, 4, 5-trisphosphate phosphatase in bovine thymus." Eur.J.Biochem.238. 350-356 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Shibata Hideki, Mukai Hideyuki, Inagaki Yoshimasa, Homma Yoshimi, Kimura Kazushi, Kaibuchi Kozo, Narumiya Shuh, Ono Yoshitaka: "Characterization of the interaction between RhoA and the amino-terminal region of PKN." FEBS Lett.385. 214-220 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Homma Miwako K., Yamasaki Motoo, Ohmi Shinobu, Homma Yoshimi.: "Inhibition of phosphoinositide hydrolysis and cell growth of Swiss 3T3 cells by myristoylated phospholipase C inhibitor peptides." J.Biochem.122. 738-742 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Homma Miwako, Homma Yoshimi.: "Anti-cancer activity of Phospholipase C inhibitor peptide." Japanese J.Cancer Chemotherapy (Japanese). 24. 165-163 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kabuyama Yukihito, Homma Yoshimi: "Phospholipid metabolism." Chemical Industry (Japanese). 48. 361-370 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Homma Yoshimi, Homma Miwako: "Phospholipase C inhibitors." Japanese J.Cancer Chemotherapy (Japanese). 24. 1611-1617 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kabuyama Yukihito, Hamaya Mikiko, Homma Yoshimi: "Wavelength specific activation of P13-kinase by UVB irradiation." FEBS Lett.441. 297-301 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Sekimata Masayuki, Emori Yasufumi, Homma Yoshimi: "Morphological changes and detachment of adherent cells induced by p122." J.Biol.chem.(in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Homma Yoshimi, Emori Yasufumi: Purification and assay of PLC-delta. In "Signalling by lnositides - A Practical Approach" (ed.SHEARS Stephen). IRL Press Oxford, 99-116 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Honma,M.K.et al.: "Inhibition of phosphoinositide hydrolysis and cell growth of Swiss 3T3 cells by myristoylated phospholipase C inhibitor peptides." J.Biochem.122(4). 738-742 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Oyama,N.et al.: "Different growth properties in response to epidermal growth factor and interleukin-6 of primary keratinocytes derived from normal and psoriatic lesional skin." J.Dermatol.Sci.16(2). 120-128 (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] 無山由己人 他: "リン脂質代謝機構" 化学と工業. 48(5). 361-370 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 本間 好 他: "ホスホリパーゼC阻害剤" 癌と化学療法. 24(11). 1611-1617 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Honma,Y.& Emori,Y.: "Signalling by Inositides;A Practical Approach." Stephen Shears edited,IRL Press, 299 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Homma,M.K.,et al.: "Growth inhibition by phospholopase C inhibitor peptides of colorectal carcinoma cells derived from familial Adenometous Polyposis." Cell Growth & Differentiation. 7. 281-288 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Qin,S.,et al.: "Cooperation of tyrosine kinases p72syk and p53/561yn regulates calcium mobilization in chicken B cell oxidant stress signaling." Eur.J.Biochem.236. 443-449 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Shibata,H.,et al.: "Characterization of the interaction between RhoA and the amino-acid region of PKN." FEBS Lett.385. 221-224 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Kabuyama,Y.,et al.: "Purification and characterization of the phosphatidylinositol-3,4,5-trisphosphate phosphatese in bovine thymus." Eur.J.Biochem.238. 350-356 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 本間 美和子、本間 好: "ホスホリパーゼC阻害ペプチドの抗癌作用" 癌と化学療法. 24(2). 323-330 (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Homma,Y.& Emori,Y.: "Purification and assay of PLC-delta" Inostol Signalling : A Practical Approach. (in press). (1997)

    • Related Report
      1996 Annual Research Report

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Published: 1996-04-01   Modified: 2016-04-21  

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