1994 年度実績報告書

細胞接着と細胞増殖におけるC-キナーゼの役割

研究課題

研究課題/領域番号	06283222
研究機関	横浜市立大学
研究代表者	大野茂男横浜市立大学, 医学部, 教授 (10142027)
研究分担者	長田真一横浜市立大学, 医学部, 助手 (00244484) 水野恵子横浜市立大学, 医学部, 助手 (90221803) 平井秀一横浜市立大学, 医学部, 講師 (80228759)
キーワード	プロテインキナーゼC / PKC関連酵素 / ホルボールエステル / 転写活性化 / ジアシルグリセロール
研究概要	C-キナーゼ(PKC)がPDGF受容体刺激に際して活性化していることを示し、これに関わるシグナル経路をPDGF受容体の自己リン酸化部位の変異体を用いて解析した。Y1021はPLCγを活性化するがP13-kinase(PI3K)その他の経路の活性化能を失っている。Y40/51はPI3Kを活性化するがPLCγ等の経路を活性化しない。F5は既知のいずれかの経路も活性化しない。nPKCεは、F5以外の変異体で活性化され、特にY40/51による活性化はPI3Kの阻害剤で完全に阻害された。これらの結果は、TPA,DG応答性の分子種、nPKCεがPLCγを介して活性化されることを細胞レベルで始めて証明したと同時に、nPKCεがPLCγとは独立に、PI3Kを介する新規の経路で活性化されることを示したこととなる。更に、化学合成したPIP3が試験管内でnPKCεを活性化し、PI3KのnPKCεに対する活性化作用がPI3Pを介したものであることも明かとなった。昨年度、同様の方法でaPKCλがPI3K経路でのみ活性化される事を示したが、この結果と合わせると従来標的の不明であったPI3Kは、TPA,DG不応答性のPKC(aPKC)と同時にTPA,DG応答性のPKC(nPKC)の直接的な上流因子であることが明かとなった。同時に従来PLC経路で活性化されるとされてきたPKCは少なくともnPKCεに関してはPLCとPI3Kにより、独立に活性化されることが明かとなった。一方、DG,PI3P,TPA等の作用部位と思われるPKCのcystein-rich domain(PKCαCRD2)の立体構造を決定した。

研究成果

(20件)

すべてその他

すべて文献書誌 (20件)

[文献書誌] Ichikawa,S,Hatanaka,H.Takeuchi,Y.,Ohno,S and Inagaki,F.: "Solution structure of cysteine-rich domain of protein kinase Cα." J.Biochem.(in press ).
[文献書誌] Mizuno,K.Noda,K.,Ueda,Y.,Hanaki,H.,Saido,T-C.,Suzuki,K.,Ikuta,T.,Kuroki,T.,Tamaoki,T.,Hirai,S-I.,Osada,S.and Ohno,S.: "UCN-01,an anti-tumor drug,is a selective inhibitor of the conventional PKC subfamily." FEBS Lett.359. 259-261 (1995)
[文献書誌] Hirano,M.,Hirai,S-I.,Mizuno,K.,Osada,S.,Hosaka,M.Ohno,S.: "Aprotein kinase Cisozyme,nPKCε,is involved in the activation of NF-κ B by 12-0-tetradecanoylphorbol-13-acetate(TPA)in rat 3Y1 fibroblasts." Biochem.Biophys.Res.Commun.206. 429-436 (1995)
[文献書誌] Kim.H-M.,Hirota,S.,Chung,H-T.,Ohno,S.,Osada,S.,Shin,T.,Ko,K-I.,Kim,J-B.,Kitamura,Y.and Nomura,S.: "Differential expression of protein kinase Cgenes in cultured mast cells derived from normal and mast-cell-deficient mice and mast cell lines." Int.Arch.Allergy Immunol.105. 258-263 (1994)
[文献書誌] Kim,H-M.,Hirota,S.,Chung,H-T.,Onouc.II.,Ito,A.,Morii,E.,Hirata,T.,Ohno,S.,Osada,S.,Kitamura,Y.and Nomura,S.: "PKCγ gene expression is dclayedin postnatal ccntral nervous system ofmi/mi mice." J.Molecular Neuroscience.4(4). 245-254 (1994)
[文献書誌] Ikuta,T.,Chida,K.,Tajima,O.,Matsuura,Y.,Iwamori,M.,Ueda,Y.,Mizuno,K.,Ohno,S.and Kuroki.T.: "Cholesterol sulfate,a novel activator for the η isoform of protein kinaseC." Cell Growth Differentiation. 5. 934-947 (1994)
[文献書誌] Fujise,A.,Mizuno,K.,Ueda,Y.,Osada,S.,Hirai,S-I.,Takayanagi,A.,Shimizu,N.,Ohwada,K.,Nakajima,H.and Ohno,S.: "Specificity of the high affinity interaction of protein kinase C witha physiological substrate,myristoylated alanine-rich C kinase substrate,MARCKS." J.Biol.Chem.269. 31642-31648 (1994)
[文献書誌] Kindregan,H.C.,Rosenbaum,S.E.,Ohno,S.and Niles,R.M.: "Characterization of conventional protein kinase C(PKC)isotype expression during F9 teratocarcinoma differentiation:Over-expression of PKC alpha alters the expression of some differentiation-dependent genes." J.Biol.Chem.269. 27756-27761 (1994)
[文献書誌] Canzian,F.,Gariboldi,M,.Manenti,G.,Gregorio,I,.Osada,S.,Ohno,S.,Dragani,T-A.and Pierotti,M-A.: "Expression in lung tumors and genetic mapping of the novel murine protein kinaseCη." Molecular Careinogenesis. 9. 111-113 (1994)
[文献書誌] Kawabe,J-I.,Iwami,G.,Ohno,S.,Katada,T.,Ueda,Y.,Homcy.C.J.and Ishikawa,Y.: "Differential activation of adenylylcyclase by protein kinase C isoenzymes" J.Biol.Chem.269. 16554-16558 (1994)
[文献書誌] Ohno,S.,Mizuno,K.,Adachi,Y.,Hata,A.,A.Y.,Akimoto,K.,Osada,S.,Hirai,S-I.and Suzuki,: "Activation of nPKCδ and ε upon mitogenic stimulation of quiescent rat3Y1 fibroblasts." J.Biol.Chem.269. 17495-17501 (1994)
[文献書誌] Mishima,K.,Ohn,S.,Shitara,N.,Yamaoka,K.and Suzuki,K.: "Opposite effects of the overexpression of protein kinaseCγ and δ on the growth properties of human glioma cell line U251MG." Biochem.Biophys.Res.Commun.201. 363-372 (1994)
[文献書誌] Ohno,S.and Suzuki,K.: "YKR2PK(S.cerevisiae)." The protein kinase factbook,Academic Press.Limited.(in press ).
[文献書誌] Ohno,S.and Suzuki,K.: "Protein KinaseC(vertebrates)Ca^<2+>-& phospholipid-dependent protein kinase." The protein kinase Factbook,Academic Press Limlted.(in press ).
[文献書誌] Hirai,S-I.,Izumi.Y.,Higa,K.,Mizuno,K.,Osada,S.,Kaibuchi,K.,Suzuki,K.and Ohno,S.: "Ras-dependent signal transduction is indispensable but not sufficientfor the activation of AP1/Jun by PKCδ." EMBOJ.13. 2331-2340 (1994)
[文献書誌] Akimoto,K.,Mizuno,K.,Osada,S.,Hirai,S-I.,Tanuma,S.,Suzuki.K.and Ohno,S.: "A new member of the third classof the protein kinaseC family,PKCλ,ubiquitously expressed dominantly in an undifferentiated mouse embyonal carcinoma cell line in and also many tissues and cells." J.Biol.Chem.269. 12677-12683 (1994)
[文献書誌] Chida,K.,Sagara,H.,Suzuki.Y.,Murakami,A.,Osada,S.,Ohno,S.,Hirosawa,K.and Kuroki,T.: "The η isoform of protein kinaseC(nPKCη)is localized on rough endoplasmic reticulum." Molecular and cellular Biol.14. 3782-3790 (1994)
[文献書誌] Koyama,H.,Inaba,M.,Nishizawa,Y.,Ohno,S.and Mori,H.: "Protein kinase C is involved in24-hydroxylase gene expression inducedby 1,25(OH)_2D_3 in rat intestinal epithelial cells." J.Cellular Biochem.55. 230-240 (1994)
[文献書誌] Suzuki,K.,Akita,Y.,Ohno,S.: "Structural and functional diversities of protein kinaseC family members." Recent Advances in Molecular and Biochemical Research on proteins,World Scientific Pub.Co.157-166 (1994)
[文献書誌] Akita,Y.,Ohno,S.,Yajima,Y.,Konno,Y.,Saido,T.C.,Mizuno,K.,Chida,K.,Osada,S.,Kuroki,T.,Kawashima,S.and Suzuki,K.: "Overproduction of a Ca^<2+>-independent protein kinase Cisozyme,nPKCε,increases the secretion of prolactin from thyrotropin-releasing hormone-stimulated rat pituitary CH_4C_1 cells." J.Biol.Chem.269-6. 4653-4660 (1994)

1994 年度 実績報告書

細胞接着と細胞増殖におけるC-キナーゼの役割

研究代表者

大野 茂男 横浜市立大学, 医学部, 教授 (10142027)

研究成果

[文献書誌] Ichikawa,S,Hatanaka,H.Takeuchi,Y.,Ohno,S and Inagaki,F.: "Solution structure of cysteine-rich domain of protein kinase Cα." J.Biochem.(in press ).

[文献書誌] Mizuno,K.Noda,K.,Ueda,Y.,Hanaki,H.,Saido,T-C.,Suzuki,K.,Ikuta,T.,Kuroki,T.,Tamaoki,T.,Hirai,S-I.,Osada,S.and Ohno,S.: "UCN-01,an anti-tumor drug,is a selective inhibitor of the conventional PKC subfamily." FEBS Lett.359. 259-261 (1995)

[文献書誌] Hirano,M.,Hirai,S-I.,Mizuno,K.,Osada,S.,Hosaka,M.Ohno,S.: "Aprotein kinase Cisozyme,nPKCε,is involved in the activation of NF-κ B by 12-0-tetradecanoylphorbol-13-acetate(TPA)in rat 3Y1 fibroblasts." Biochem.Biophys.Res.Commun.206. 429-436 (1995)

[文献書誌] Kim,H-M.,Hirota,S.,Chung,H-T.,Onouc.II.,Ito,A.,Morii,E.,Hirata,T.,Ohno,S.,Osada,S.,Kitamura,Y.and Nomura,S.: "PKCγ gene expression is dclayedin postnatal ccntral nervous system ofmi/mi mice." J.Molecular Neuroscience.4(4). 245-254 (1994)

[文献書誌] Ikuta,T.,Chida,K.,Tajima,O.,Matsuura,Y.,Iwamori,M.,Ueda,Y.,Mizuno,K.,Ohno,S.and Kuroki.T.: "Cholesterol sulfate,a novel activator for the η isoform of protein kinaseC." Cell Growth Differentiation. 5. 934-947 (1994)

[文献書誌] Canzian,F.,Gariboldi,M,.Manenti,G.,Gregorio,I,.Osada,S.,Ohno,S.,Dragani,T-A.and Pierotti,M-A.: "Expression in lung tumors and genetic mapping of the novel murine protein kinaseCη." Molecular Careinogenesis. 9. 111-113 (1994)

[文献書誌] Kawabe,J-I.,Iwami,G.,Ohno,S.,Katada,T.,Ueda,Y.,Homcy.C.J.and Ishikawa,Y.: "Differential activation of adenylylcyclase by protein kinase C isoenzymes" J.Biol.Chem.269. 16554-16558 (1994)

[文献書誌] Ohno,S.,Mizuno,K.,Adachi,Y.,Hata,A.,A.Y.,Akimoto,K.,Osada,S.,Hirai,S-I.and Suzuki,: "Activation of nPKCδ and ε upon mitogenic stimulation of quiescent rat3Y1 fibroblasts." J.Biol.Chem.269. 17495-17501 (1994)

[文献書誌] Mishima,K.,Ohn,S.,Shitara,N.,Yamaoka,K.and Suzuki,K.: "Opposite effects of the overexpression of protein kinaseCγ and δ on the growth properties of human glioma cell line U251MG." Biochem.Biophys.Res.Commun.201. 363-372 (1994)

[文献書誌] Ohno,S.and Suzuki,K.: "YKR2PK(S.cerevisiae)." The protein kinase factbook,Academic Press.Limited.(in press ).

[文献書誌] Ohno,S.and Suzuki,K.: "Protein KinaseC(vertebrates)Ca^<2+>-& phospholipid-dependent protein kinase." The protein kinase Factbook,Academic Press Limlted.(in press ).

[文献書誌] Hirai,S-I.,Izumi.Y.,Higa,K.,Mizuno,K.,Osada,S.,Kaibuchi,K.,Suzuki,K.and Ohno,S.: "Ras-dependent signal transduction is indispensable but not sufficientfor the activation of AP1/Jun by PKCδ." EMBOJ.13. 2331-2340 (1994)

[文献書誌] Chida,K.,Sagara,H.,Suzuki.Y.,Murakami,A.,Osada,S.,Ohno,S.,Hirosawa,K.and Kuroki,T.: "The η isoform of protein kinaseC(nPKCη)is localized on rough endoplasmic reticulum." Molecular and cellular Biol.14. 3782-3790 (1994)

[文献書誌] Koyama,H.,Inaba,M.,Nishizawa,Y.,Ohno,S.and Mori,H.: "Protein kinase C is involved in24-hydroxylase gene expression inducedby 1,25(OH)_2D_3 in rat intestinal epithelial cells." J.Cellular Biochem.55. 230-240 (1994)

[文献書誌] Suzuki,K.,Akita,Y.,Ohno,S.: "Structural and functional diversities of protein kinaseC family members." Recent Advances in Molecular and Biochemical Research on proteins,World Scientific Pub.Co.157-166 (1994)

1994 年度実績報告書

大野茂男横浜市立大学, 医学部, 教授 (10142027)