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Involvement of phosphatidylinositol 3-kinase in cell growth

Research Project

Project/Area Number 03454163
Research Category

Grant-in-Aid for General Scientific Research (B)

Allocation TypeSingle-year Grants
Research Field Pathological medical chemistry
Research InstitutionThe University of Tokyo (1992)
Tokyo Metropolitan Institute of Gerontology (1991)

Principal Investigator

TAKENAWA Tadaomi  Institute of Medical Science, Univ. of Tokyo, Professor, 医科学研究所, 教授 (40101315)

Co-Investigator(Kenkyū-buntansha) SHIBASAKI Futoshi  Tokyo Metropolitan Institute of Gerontology, Research associate, 生体情報研究部門, 研究員 (40241260)
山川 彰夫  東京都老人総合研究所, 研究員 (30200588)
Project Period (FY) 1991 – 1992
Project Status Completed (Fiscal Year 1992)
Budget Amount *help
¥7,100,000 (Direct Cost: ¥7,100,000)
Fiscal Year 1992: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1991: ¥5,600,000 (Direct Cost: ¥5,600,000)
KeywordsPI3 Kinase / Tyrosine kinase / Purification / SH_2 / Tyrosine phosphorylation / PI3キナ-ゼ / SH_3 / αーアクチニン / 細胞骨格系
Research Abstract

Two types of phosphatidylinositol(PI) 3-kinase (PI3K) have been purified 6250-fold (PI3KI) and 1250-fold (PI3KII) from the cytosol fraction of bovine thymus, respectively. Purified PI3KI and PI3KII were found to be apparent molecular weight of 110 KDa and 190 KDa respectively by a gel filtration. On the other hand, on SDS gel electrophoresis molecular weight of PI3KI was also estimated as 110 KDa but PI3KII showed two bands of which molecular weight are 110 KDa and 85 KDa, suggesting a heterodimer form. Peptide mapping analysis also demonstrated that a 110 KDa protein contained in PI3KII is the same protein as PI3KI. Specific activity of PI3KI was calculated as 250 nmol/min/mg protein, while that of PI3KII was as 25 nmol/min/mg protein, indicating that monomer has higher activity than heterodimer. PI3KII but not PI3KI was co-immunoprecipitated with pp60^<v-src> and middle T/pp60^<c-src> even in the conditions where PI3Ks were not phosphorylated suggesting that non phosphorylated PI3K recognized pp60^<v-src> PI3KII was phosphorylated by pp60^<v-src> and bound to pp60^<v-src>. Anti-p85 (85 Kda subunit of PI3KII) antibody which precipitated PI3KII co-immunoprecipitated pp60^<v-src> in src-transformed cells suggesting that PI3KII bound to pp60^<v-src>. All these data shows that these PI3Ks are regulated independently.

Report

(3 results)
  • 1992 Annual Research Report   Final Research Report Summary
  • 1991 Annual Research Report
  • Research Products

    (36 results)

All Other

All Publications (36 results)

  • [Publications] H.Kato et al.: "Activation of phospholipase C delta in the aortas of spontaneously hypertensive rats" J.Biol.Chem.267. 6483-6487 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Y.Homma et al.: "Activation of phospholipase C as well as tyrosine kinase in rat 3Y1 fibroblast by expression of srcregion of phospholipase C-δ2" J.Biol.Chem.267. 3778-3782 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Y.HOmma et al.: "Purification of recobinant SH2/SH3 proteins of phospholipase Cδ1 and δ2 and their inhibitory effect on PIP2 hydrolysis induced by both type of phospholipase Cδ" Biochem.Biophys.Res.Commun.182. 1402-1407 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] K.Shimazaki et al.: "Purification of AMPA type glutamate receptor by a spider toxin" Mol.Brain Res.13. 331-337 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] K.Furuhashi et al.: "Inositol phospholypid-induced suppression of F-actingelating activity of smooth muscle filamin" Biochem.Biophys.Res.Commun.184. 1261-1265 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] K.Fukami et al.: "Phosphatidic acid that accumulates in PDGF-stimulated Balb/c 3T3 cells in a potential mitogenic signal" J.Biol.Chem.267. 10988-10993 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] K.Fukami et al.: "Requierment of phosphatidylinositol 4,5-bisphosphate for -actinin function" Nature. 359. 150-152 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] K.Matsuoka et al.: "Cloning of Ash, a ubiquitous protein composed of SH2 and SH3 domains in human and rat" Proc.Natl.Acad.Sci.USA. 89. 9015-9019 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] K.Shiroki et al.: "Adenovirus EIA proteins stimulate the inositol phospholipid metabolism in PCI2 cells" J.Virology. 66. 6093-6098 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] F.Shibasaki et al.: "Properties of phosphatidylinositol 3-kinase in bovine thymus" Biochem.J.289. 227-231 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] N.Kawai et al.: "Neuroreceptors,Ion-channels and the Brain" Elsevier Science Publishers, (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] H.Kato, F.Shibasaki and T.Takenawa: "Activation of phospholipase C delta in the aortas of spontaneously hypertensive" J. Biol. Chem. 267. 6483-6487 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Y.Homma, Y.Emori and T.Takenawa: "Activation of phospholipase C as well as tyrosine kinase in rat 3Y1 fibroblast by expression of src-homology region of phospholipase c-g2" J. Biol. Chem. 267. 3778-3782 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Y.Homma, Y.Emori, and T.Takenawa: "Purification of recombinant SH2/SH3 proteins of phospholipase Cg1and g2 and their inhibitory effect on PIP2 hydrolysis induced by both types of phospholipase Cg" Biochem. Biophys. Res. Commun. 182. 1402-1407 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] K.Shimazaki, P.C.Hugh, T.Nakajima, N.Kawai and T.Takenawa: "Purification of AMPA type glutamate receptor by a spider toxin" Mol. Brain Res. 13. 331-337 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] N.Kawai, A.Miwa, K.Shimazaki, H.C.P.Robinson, T.Takenawa and T.Nakajima: "Glutamate receptors and neurotoxinsin Neuroreceptors" Ion channels and the Brain (ed. by N.Kawai) Elsevier Science Publishers. p3-10 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] K.Furuhashi, M.Inagaki, S.Hatano, K.Fukami and T.Takenawa: "Inositol phospholipid-induced suppression of Factin-gelating activity of smooth muscle filamin" Biochem. Biophys. Res. Commun. 184. 1261-1265 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] K.Fukami and T.Takenawa: "Phosphatidic acid that accumulates in PDGF-stimulated Balb/c 3T3 cells in a potential mitogenic signal" J. Biol. Chem. 267. 10988-10993 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] K.Fukami, K.Furuhashi, M.Inagaki, T.Endo, S.Hatano and T.Takenawa: "Requirement of phosphatidylinositol 4,5-bisphosphate for a-actinin function" Nature. 359. 150-152 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] K.Matsuoka, M.Shibata, A.Yamakawa and T.Takenawa: "Cloning of Ash, a ubiquitous protein composed of SH2 and SH3 domains in human and rat" Proc. Natl. Acad. Sci. USA. 89. 9015-9019 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] K.Shiroki, A.Yamakawa, M.Shibata, T.Takenawa and A.Nomoto: "Adenovirus E1A proteins stimulate the inositol phospholipid metabolism in PC12 cells" J. Virology. 66. 6093-6098 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] F.Shibasaki, Y.Fukui and T.Takenawa: "Properties of phosphatidylinositol 3-kinase in bovine thymus" Biochem. J.289. 227-231 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] H.Kato et al.: "Activation of phospholipase C delta in the aortas of spontaneously hypertensive rats" J.Biol.Chem.267. 6483-6487 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Y.Homma et al.: "Activation of phospholipase C as well as tyrosine kinase in rat 3Y1 fibroblast by expression of srcregion of phospholipase C-X2" J.Biol.Chem.267. 3778-3782 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Y.Homma et al.: "Purification of recombinant SH2/SH3 proteins of phospholipase Cδ1 and δ2 and their inhibitory effect on PIP2 hydrolysis induced by both type of phospholipase Cδ" Biochem.Biophys.Res.Commum.182. 1402-1407 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] K.Shimazaki et al.: "Purification of AMPA type glutamate receptor by a spider toxin" Mol.Brain Res.13. 331-337 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] K.Furuhashi et al.: "Inositol Phospholypid-induced suppression of F-actin-gelating activity of smooth muscle filamin" Biochem.Biophys.Res.Commun.184. 1261-1265 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] K.Fukami et al.: "Phosphatidic acid that accimulates in PDGF-stimulated Ba1b/c 3T3 cells in a potential mitogenic signal" J.Biol.Chem.267. 10988-10993 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] K.Fukami T.Takenawa et al.: "Requierment of phosphatidylinositol 4,5-bisphosphate for δ-actinin function" Nature. 359. 150-152 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] K.Matsuoka,T.Takenawa et al.: "Cloning of Ash, a ubiquitous protein composed of SH2 and SH3 domains in human and rat" Proc.Natl.Acad.Sci.USA. 89. 9015-9019 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] K.Shiroki,T.takenawa et al.: "Adenovirus EIA proteins stimulate the inositol phospholipid metabllism in PCI2 cells" J.Virology. 66. 6093-6098 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] F.Shibasaki T.Takenawa: "Properties of phosphatidylinositol 3-kinase in bivine thymus" Biochem.J.289. 227-231 (1993)

    • Related Report
      1992 Annual Research Report
  • [Publications] N.Kawai et al.: "Neuroreceptors,Ton-channels and the Brain" Elsevier Science Publishers, (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Hidaka,M.: "Highly conserved eight amino acid sequence in SH_2 is important for recognition of phosphotyrosine site" Biochem.Biophys.Res.Commun.180. 1490-1497 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Shibasaki,F.: "Two types of phosphatidylinositol kinase from bovine thymus" J.Biol.Chem.266. 8108-8111 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Hayashi,H,: "Phosphorylation in vitro of 85KDa subunit of phosphatidylinositol 3-kinase and its possible activation by insulin receptor tyrosine kinase" Biochem J.

    • Related Report
      1991 Annual Research Report

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Published: 1991-04-01   Modified: 2016-04-21  

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