1992 Fiscal Year Annual Research Report

蛋白質チロシン残基の燐酸化とその細胞生物学的意義

Research Project

Project/Area Number	02304034
Research Institution	Fukui Medical School
Principal Investigator	山村博平福井医科大学, 医学部, 教授 (90030882)
Co-Investigator(Kenkyū-buntansha)	山本雅東京大学, 医科研, 教授 (40134621) 蛯名洋介徳島大学, 酵素研, 教授 (00112227) 武田誠郎広島大学, 医学部, 教授 (40030853) 春日雅人神戸大学, 医学部, 教授 (50161047) 中川八郎大阪大学, 蛋白研, 教授 (20029937)
Keywords	CDNAクローニング / CSK / p60^<src> / インスリン受容体 / PI-3キナーゼ / EGF受容体チロシンキナーゼ / P72^<syk> / 蛋白質チロシンホスファターゼ
Research Abstract	1)src familyチロシンキナーゼに関して、pp60^<c-src>を燐酸化するc-src kinase(CSK)の細胞機能における役割を明かにするため、CSK遺伝子を欠損させた動物の作成したところ、CSK欠損個体は胎生11日目で到死となった。その胚を検索すると神経管の発達不全、頭部神経冠細胞の細胞死が確認された。またこれらの組織においてpp60^<c-src>、pp59^<fyn>、po53/56^<lyn>の顕著な活性化が認められた。これらの結果はsrc familyは神経系の発生過程に何らかの重要な働きをしていることを示唆している。またp60^<c-yes>、pp59^<fyn>の自己燐酸化による活性化について検討し分子間反応であることを明かにした。さらに正常細胞における他のsrc familyチロシンキナーゼに関してfynやlynの標的蛋白質について検討した結果、lynの下流にはp75^<HS1>が存在すること、fynの下流にはVavやPLC-γ、MAPキナーゼ等の存在を示唆することが出来た。新しく見いだされた非受容体型チロシンキナーゼであるp72sykは血小板においてpp60^<c-src>の約半量存在し、生理的なリガンドであるスロンビンによって活性化されるがその活性化解除においてはCa^<2+>の存在が必要である。このCa^<2+>の作用は阻害剤の実験からミオシン軽鎖キナーゼが関与している可能性が強い。2)増殖因子受容体チロシンキナーゼにおいてはインスリンで細胞を刺激するとインスリン受容体キナーゼ依存性にイノシトール燐脂質3キナーゼの85Kサブユニットのチロシン残基が燐酸化されることを明かにした。さらにその燐酸化部位は368,580,607番目のチロシン残基である。またインスリン受容体キナーゼはIRS(Insulin-receptor substrate-1)を燐酸化しその燐酸化部位をイノシトール燐脂質3キナーゼの85KのSH2の領域が認識する可能性を示唆した。さらに上皮成長因子受容体チロシンキナーゼの下流にG蛋白質に関係するGAP(GTPase activating protein)が存在し、この系が細胞の増殖分化において何らかの重要な役割を演じていることを示唆した。3)チロシンホスファターゼに関して肝臓において4種類(L1-4)分離同定することが出来た。そのうちL1は2つのSH2領域をもつ全く新しいホスファターゼであることを明かにした。

Research Products
(34 results)

All Other

All Publications (34 results)

[Publications] Iwashita,S.: "Signal transduction system for growth factor receptors associated with tyrosine kinase activity:Epidermal growth factor receptor signalling its regulation." Cellular Signalling,. 4. 123-132 (1992)
[Publications] Nakamura,T.: "Isolation and characterization of activin receptor from mouse embryonal carcinoma cells.Identification of its serine/threonine/tyrosine protein kinase activity." J.Biol.Chem.267. 18924-18928 (1992)
[Publications] Sabe,H.: "Molecular cloning and expression of chicken C-terminal Src kinase:lack of stable association with c-Src protein." Proc.Natl.Acad.Sci.USA. 89. 2190-2194 (1992)
[Publications] Sabe,H.: "Activation of c-Src in cells bearing v-Crk and its suppression by Csk." Mol.Cell.Biol.12. 4706-4713 (1992)
[Publications] Junichiro Muzuguchi: "Tyrosine Protein kinase is involved anti-IgM-mediated signalling in Ball7 B lymphoma cells." J.Immunol.148. 689-694 (1992)
[Publications] Yuji.Yamanashi: "Activation of Src-like protein-tyrosine kinase Lyn and Lyn associated PI-3 kinase upon B cell antigen receptor-mediated signaling." Proc.Natl.Acad.Sci.USA. 89. 1118-1122 (1992)
[Publications] Koko Katagiri: "Modulation of monocytic differentiation of HL60 cells by inhibitors of protein-tyrosine kinases." Cell Immunol.140. 282-294 (1992)
[Publications] Hisashi Umemori: "Specific expression of antigen receptor-associated tyrosine kinases,Fyn and Lyn,in the central nernous system." Mol.Cell Biol.12. 3784-3795 (1992)
[Publications] Fumiaki Uchiumi: "Characterization of the promoter region of the src-family gene lyn and its trans-activation by HTLV-I encoded p40tax." Mol.Cell.Biol.12. 3784-3795 (1992)
[Publications] Takahiro Miyauchi: "Purification and characterization of a possible protooncogene fyn product,p59^<fyn>,from a rat brain particulate fraction." J.Biochem.112. 729-732 (1992)
[Publications] Takeuchi,F.: "Phosphorylation of histone H2A by protein kinase C and identification of the phosphorylation sete." J.Biochem.111(6). 788-792 (1992)
[Publications] Ohta,S.: "Protein-tyrosine phosphorylations induced by concanavalin A and N-Formyl-methionyl-phenylalanine in human neutrophils." Eur.J.Biochem.206(3). 895-900 (1992)
[Publications] Ohta,S.: "Protein-tyrosine kinase p72^<syk> is activated by wheat germ agglutinin in platelets." Biochem.Biophys.Res.commun.185(3). 1128-1132 (1992)
[Publications] Asahi,M.: "Thrombin-induced human platelet aggregation is inhibited by protein-tyrosine kinase inhibitors,ST638 and genistein." FEBS Lett.309(1). 10-14 (1992)
[Publications] Takeuchi,N.: "Inhibotory effect of mitoxantrone on activity of protein kinase C and Growth of HL60 cells." J.Bichem.112(6). 762-767 (1992)
[Publications] Hayashi,K.: "Genomic structure of human caldesmon gene." Proc.Natl.Acad.Sci.USA. 89. 12116-12122 (1992)
[Publications] Yonezawa,K.: "In vitro association of phosphatidylinositol 3-kinase activity with the activated insulin receptor tyrosine kinase." J.Biol.Chem.267. 440-446 (1992)
[Publications] Chatani,Y.: "Mitogen-induced tyrosine-phosphorylated 41-and 43-kDa proteins are family members of extracellular signal-regulated kinases/microtubule-associated protein 2 Kinases." J.Biol.Chem.267. 9911-9916 (1992)
[Publications] Ando,A.: "Enhanced insulin-induced mitogenesis and mitogen-activated protein kinase activities in mutant insulin receptors with substitution two COOH-terminal tyrosine autophosphorylation sites by phenylalanine." J.Biol.Chem.267. 12788-12796 (1992)
[Publications] Yonezawa,K.: "Insulin-dependent formation of a complex containing 85-kDa subunit of PI3-kinase and tyrosine-phosphorylated IRS-1." J.Biol.Chem.267. 25958-25966 (1992)
[Publications] Takashi Murakami: "Idenification of Two Enhancer Elements in the Gene Encoding the Type 1 Glucose Transporter from the Mouse Which Are Responsive to Serum,Growth Factor,and Oncogenes." J.Biol.Chem.267. 9300-9306 (1992)
[Publications] Hideki Hayashi: "Insulin Treatment Stimulates the Tyrosine Phosphorylation of the α-Type 85-kDa Subunit of Phosphatidylinositol 3-Kinase in Vivo." J.Biol.Chem.267. 22575-22580 (1992)
[Publications] Shimada,F.: "Abnormal messenger ribonucleic acid(mRNA)transcribed from a mutant insulin receptor gene in a patient with type A insulin resistance." Diabetologia. 35. 639-644 (1992)
[Publications] Makino,H.: "Insulin receptor gene mutation:a molecular genetical and functional analysis." Cellular Signaling. 4. 351-363 (1992)
[Publications] Akira Hiraga: "Purification and characterization of a rat liver protein-tyrosine phosphatase with sequence similarity to src-homology region 2." Eur.J.Biochem.209(1). 195-206 (1992)
[Publications] Taniguchi,T.: "Protein-tyrosine kinase p72^<syk> is axtivated by thrombin in platelets." J.Biol.Chem.268(4). 2277-2279 (1993)
[Publications] Akira Hiraga: "Identification of a rat liver protein-tyrsine phosphatase similar to human placental PTPase-1B using quantitatively phosphorylated protein substrates." J.Biochem.113(2). 180-188 (1993)
[Publications] Marius Sudol: "Novel Yes-related kinase,Yrk,is expressed at elevated levels in neural and hematopoietic tissues." Oncogene.
[Publications] Tanaka,J.: "Morpholohical and Biochemical analyses of contractile proteins (actin,myosin,coldesmon and tropomyosin)in normal and transformed cells." J.Cell Sci.
[Publications] Hideki Hayashi: "The a-Type 85-kDa Subunit of Phosphatidylinositol 3-Kinase Phosphorylated at Tyrosines368,580,and 607 by the Insulin Receptor." J.Biol.Chem.
[Publications] Yuji Yamanashi: "Identificarion of HS1 protein as a major substrate of protein-tyrosine kinase(s) upon B cell antigen receptor-mediated signaling." Proc.Natl.Acad.Sci.USA.
[Publications] Takahiro Miyauchi: "Purification and characteriation of the possible protooncogene fyn product,p59fyn,from rat brain particulate fraction." J.Biochem.
[Publications] Koko Katagiri: "Tyrosine phosphorylation of tubulins during monocytic differentiation of HL-60 cells." J.Immunol.
[Publications] Tadashi Yamamoto: "Association of Src-Family kinase Lyn with B-cell antigen receptor." Immu.Rev.

1992 Fiscal Year Annual Research Report

蛋白質チロシン残基の燐酸化とその細胞生物学的意義

Principal Investigator

山村 博平 福井医科大学, 医学部, 教授 (90030882)

Research Products

[Publications] Iwashita,S.: "Signal transduction system for growth factor receptors associated with tyrosine kinase activity:Epidermal growth factor receptor signalling its regulation." Cellular Signalling,. 4. 123-132 (1992)

[Publications] Nakamura,T.: "Isolation and characterization of activin receptor from mouse embryonal carcinoma cells.Identification of its serine/threonine/tyrosine protein kinase activity." J.Biol.Chem.267. 18924-18928 (1992)

[Publications] Sabe,H.: "Molecular cloning and expression of chicken C-terminal Src kinase:lack of stable association with c-Src protein." Proc.Natl.Acad.Sci.USA. 89. 2190-2194 (1992)

[Publications] Sabe,H.: "Activation of c-Src in cells bearing v-Crk and its suppression by Csk." Mol.Cell.Biol.12. 4706-4713 (1992)

[Publications] Junichiro Muzuguchi: "Tyrosine Protein kinase is involved anti-IgM-mediated signalling in Ball7 B lymphoma cells." J.Immunol.148. 689-694 (1992)

[Publications] Yuji.Yamanashi: "Activation of Src-like protein-tyrosine kinase Lyn and Lyn associated PI-3 kinase upon B cell antigen receptor-mediated signaling." Proc.Natl.Acad.Sci.USA. 89. 1118-1122 (1992)

[Publications] Koko Katagiri: "Modulation of monocytic differentiation of HL60 cells by inhibitors of protein-tyrosine kinases." Cell Immunol.140. 282-294 (1992)

[Publications] Hisashi Umemori: "Specific expression of antigen receptor-associated tyrosine kinases,Fyn and Lyn,in the central nernous system." Mol.Cell Biol.12. 3784-3795 (1992)

[Publications] Fumiaki Uchiumi: "Characterization of the promoter region of the src-family gene lyn and its trans-activation by HTLV-I encoded p40tax." Mol.Cell.Biol.12. 3784-3795 (1992)

[Publications] Takahiro Miyauchi: "Purification and characterization of a possible protooncogene fyn product,p59^<fyn>,from a rat brain particulate fraction." J.Biochem.112. 729-732 (1992)

[Publications] Takeuchi,F.: "Phosphorylation of histone H2A by protein kinase C and identification of the phosphorylation sete." J.Biochem.111(6). 788-792 (1992)

[Publications] Ohta,S.: "Protein-tyrosine phosphorylations induced by concanavalin A and N-Formyl-methionyl-phenylalanine in human neutrophils." Eur.J.Biochem.206(3). 895-900 (1992)

[Publications] Ohta,S.: "Protein-tyrosine kinase p72^<syk> is activated by wheat germ agglutinin in platelets." Biochem.Biophys.Res.commun.185(3). 1128-1132 (1992)

[Publications] Asahi,M.: "Thrombin-induced human platelet aggregation is inhibited by protein-tyrosine kinase inhibitors,ST638 and genistein." FEBS Lett.309(1). 10-14 (1992)

[Publications] Takeuchi,N.: "Inhibotory effect of mitoxantrone on activity of protein kinase C and Growth of HL60 cells." J.Bichem.112(6). 762-767 (1992)

[Publications] Hayashi,K.: "Genomic structure of human caldesmon gene." Proc.Natl.Acad.Sci.USA. 89. 12116-12122 (1992)

[Publications] Yonezawa,K.: "In vitro association of phosphatidylinositol 3-kinase activity with the activated insulin receptor tyrosine kinase." J.Biol.Chem.267. 440-446 (1992)

[Publications] Chatani,Y.: "Mitogen-induced tyrosine-phosphorylated 41-and 43-kDa proteins are family members of extracellular signal-regulated kinases/microtubule-associated protein 2 Kinases." J.Biol.Chem.267. 9911-9916 (1992)

[Publications] Ando,A.: "Enhanced insulin-induced mitogenesis and mitogen-activated protein kinase activities in mutant insulin receptors with substitution two COOH-terminal tyrosine autophosphorylation sites by phenylalanine." J.Biol.Chem.267. 12788-12796 (1992)

[Publications] Yonezawa,K.: "Insulin-dependent formation of a complex containing 85-kDa subunit of PI3-kinase and tyrosine-phosphorylated IRS-1." J.Biol.Chem.267. 25958-25966 (1992)

[Publications] Takashi Murakami: "Idenification of Two Enhancer Elements in the Gene Encoding the Type 1 Glucose Transporter from the Mouse Which Are Responsive to Serum,Growth Factor,and Oncogenes." J.Biol.Chem.267. 9300-9306 (1992)

[Publications] Hideki Hayashi: "Insulin Treatment Stimulates the Tyrosine Phosphorylation of the α-Type 85-kDa Subunit of Phosphatidylinositol 3-Kinase in Vivo." J.Biol.Chem.267. 22575-22580 (1992)

[Publications] Shimada,F.: "Abnormal messenger ribonucleic acid(mRNA)transcribed from a mutant insulin receptor gene in a patient with type A insulin resistance." Diabetologia. 35. 639-644 (1992)

[Publications] Makino,H.: "Insulin receptor gene mutation:a molecular genetical and functional analysis." Cellular Signaling. 4. 351-363 (1992)

[Publications] Akira Hiraga: "Purification and characterization of a rat liver protein-tyrosine phosphatase with sequence similarity to src-homology region 2." Eur.J.Biochem.209(1). 195-206 (1992)

[Publications] Taniguchi,T.: "Protein-tyrosine kinase p72^<syk> is axtivated by thrombin in platelets." J.Biol.Chem.268(4). 2277-2279 (1993)

[Publications] Akira Hiraga: "Identification of a rat liver protein-tyrsine phosphatase similar to human placental PTPase-1B using quantitatively phosphorylated protein substrates." J.Biochem.113(2). 180-188 (1993)

[Publications] Marius Sudol: "Novel Yes-related kinase,Yrk,is expressed at elevated levels in neural and hematopoietic tissues." Oncogene.

[Publications] Tanaka,J.: "Morpholohical and Biochemical analyses of contractile proteins (actin,myosin,coldesmon and tropomyosin)in normal and transformed cells." J.Cell Sci.

[Publications] Hideki Hayashi: "The a-Type 85-kDa Subunit of Phosphatidylinositol 3-Kinase Phosphorylated at Tyrosines368,580,and 607 by the Insulin Receptor." J.Biol.Chem.

[Publications] Yuji Yamanashi: "Identificarion of HS1 protein as a major substrate of protein-tyrosine kinase(s) upon B cell antigen receptor-mediated signaling." Proc.Natl.Acad.Sci.USA.

[Publications] Takahiro Miyauchi: "Purification and characteriation of the possible protooncogene fyn product,p59fyn,from rat brain particulate fraction." J.Biochem.

[Publications] Koko Katagiri: "Tyrosine phosphorylation of tubulins during monocytic differentiation of HL-60 cells." J.Immunol.

[Publications] Tadashi Yamamoto: "Association of Src-Family kinase Lyn with B-cell antigen receptor." Immu.Rev.

山村博平福井医科大学, 医学部, 教授 (90030882)