1994 年度実績報告書

プロテインキナーゼC群およびその優性抑制変異体群の作用機構の解析

研究課題

研究課題/領域番号	05454162
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, 助手 (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 / キナーゼ / ジアシルグリセロール
研究概要	昨年見いだしたPKCの優性抑制変異体に加え、本年はまずPKCdeltaの活性型変異体を作成した。これらを用いて、PKC経路とrasを介する経路との相互関係を分子生物学的に調べることが始めて可能となった。PKCの変異体と活性型ras及び優性抑制型のras等を用いて、TPA→PKC→TRE(TPA応答性因子)へのシグナル経路におけるrasの作用を種々の角度から検討した。その結果PKCを介するシグナル経路においてrasは必須であるが、それだけでは充分ではないことが明かとなった。すなわち、TPA→PKC→ras→TREという経路が存在すること。さらに、これとは独立にTPA→PKC→?→TREという経路が存在することが明かとなった。ここに至ってPKCからrasに至る経路に介在する分子を同定することが大きな問題となってきた。クローン化された種々の組換え蛋白を用いた解析から、おそらくras自身はPKCと結合もしないし、PKCによって影響を受けないことが明かとなり、現在rasの活性を制御する種々の蛋白についてPKCとの関わりを解析中である。一方、PKCの未知の下流因子、すなわち基質蛋白質を見いだす目的で、PKCの優性抑制変異体に直接結合する細胞蛋白質の直接クローニンングを開始した。

研究成果

(19件)

すべてその他

すべて文献書誌 (19件)

[文献書誌] Hirai,S.,Izumi,Y.,Higa,K.,Mizuno,K.,Osada,S.,Kaibuchi,K.,Suzuki,K.and Ohno,S.: "Ras-dependent signal transducition is indispensable but not sufficient for the activation of AP1/Jun by PKCdelta." EMBOJ.(in press).
[文献書誌] Akimoto,K.,Mizuno,K.,Osada,S.,Hirai,S.,Tanuma,S.,Suzuki,K.and Ohno,S.20GB02:A new member of the third class of the protein kinase C family,PKClambda,ubiquitously expressed dominantly in an undifferentiated mouse embyonal carcinoma cell line and also in many tissues and cells.: J.Biol.Chem.(in press).
[文献書誌] Ohno,S.and Suzuki,K.: "Protein kinase C:Reassessment.Function of the novel members of the protein kinase C family." FESEB J.(in press).
[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 C isozyme,nPKC epsilon,increases the secretion of protein from thyrotropin-releasing hormone-stimulated rat pituitary GH_4C_1 cells." J.Biol.Chem.(in press).
[文献書誌] Chida,K.,Sagara,H.,Suzuki,Y.,Murakami,A.,Osada,S.,Ohno,S.,Hirosawa,K,and Kuroki,T.: "The eta isoform of protein kinase C(nPKCeta)is localized on rough endoplasmic reticulum." Molecular and Cellular Biol.(in press).
[文献書誌] Koyama,H.,Inaba,M.,Nishizawa,Y.,Ohno,S.and Morii,H.: "Protein kinase C is involved in 24-hydroxylase gene expression induced by 1,25(OH)_2D_3 in rat intestinal epithelial cells." J Cellular Biochemistry. (in press).
[文献書誌] Suzuki,K.,Akita,Y.and Ohno,S.: "Structural and functional diversities of protein kinase C family members." Recent Advances in Molecular and Biochemical Research on proteins.(in press).
[文献書誌] Ohno,S.,Akita,Y.,Hata,A.,Osada,S.,Kubo,K.,Konno,Y.,Akimoto,K.,Mizuno,K.,Saido,T.C.,Kuroki,T.and Suzuki,K.: "Structural and functional diversities of a family of signal transducing protein Kinases;protein Kinase C family." Yckohama Med.Bull.43. 139-140 (1993)
[文献書誌] Ohno,S.: "Diversity and specificity of multiple protein kinase C." Progress in Endocrinology,Parthenon Press. 63-66 (1993)
[文献書誌] Koizumi,H.,Kohno,Y.,Osada,S.,Ohno,s.,Ohkawara,and Kuroki,T.: "Differentiation-associated localization of nPKCeta,a Ca^<++>-independent protein kinase C,in normal human skin and skin diseases." J.Investigative Dermatology. 6. 858-863 (1993)
[文献書誌] Mischak,H.,Goodnight,J.,Henderson,D.W.,Osada,S.,Ohno,s.and Mushinski,F.: "Unique expression pattern of protein kinase C-theta:high _mRNA levels in normal mouse testes and in T-lymphocytic cells and neoplasms." FEBS Lett.326. 51-55 (1993)
[文献書誌] Sublette,E.,Naik,M.U.,Jiang,X,Osten,P.,Valsarmis,H.,Osada,S.,Ohno,S.and Sacktor,T.C.: "Evidence for a new,high-molecular weight isoform of protein kinase C in rat hippocampus." Neuroscience Lett.159. 175-178 (1993)
[文献書誌] Mizuno,K.,Saido,T.C.,Ohno,S.,Tamaoki,T.and Suzuki,K.: "Staurosporine-related compounds,K252a and UCN-01,inhibits both cPKC and nPKC." FEBS Lett.330. 114-116 (1993)
[文献書誌] Park,H.-Y.,Russakovsky,V.,Ohno,S.and Gilchrest,B.A.: "The beta-Isoform of protein kinase C stimulates human melanogenesis by act:rating tyrosinase in pigment cells." J.Biol.Chem.268. 11742-11749 (1993)
[文献書誌] Hata,A.,Akita,Y.,Suzuki,K.and Ohno,S.: "Functional divergence of protein kinase C(PKC)family members;PKCgamma differs from PKC alpha,betaII,and nPKCepsilon in its competence to mediate 12-O-tetradecanoyl phorbol 13-acetate(TPA)-responsive transcriptional activation through a TPA-response element." J.Biol.Chem.268. 9122-9129 (1993)
[文献書誌] Osada,S.,Hashimoto,Y.,Nomura,S.,Konnno,Y.,Cihida,K.,Tajima,O.,Kubo,K.,Akimoto,K.,Koizumi,H.,Kitamura,Y.,Suzuki,K.,Ohno,S.and Kuroki,T.: "Predominant expression of novel protein kinase C eta,aCa^<2+>-independent isoform of protein kinase C in epithelial tissues,in association with epithlial differentiation." Cell Growth & Differentiation. 4. 167-175 (1993)
[文献書誌] Wang,F.,Naik,U,P.,Ehrlich,Y,H.,Freyberg Z.,Osada,S.,Ohno,S.,Kuroki,T.,Suzuki,K,and Kamecki,E.: "A new protein kinase C,nPKC eta′ and PKC theta are expressed in human platelets : involvement of nPKCn'and nPKC theta in signal transduction stimulated by PAF." Biochem.Biophys.Res.Commun.191. 240-246 (1993)
[文献書誌] 大野茂男: "Cキナーゼファミリーの活性調節" 細胞増殖の制御,南江堂. 105-114 (1993)
[文献書誌] 大野茂男: "細胞増殖と疾患-細胞増殖の細胞内シグナル伝達機構" Mebio,メジカルビュー社. 10,6. 30-38 (1993)

1994 年度 実績報告書

プロテインキナーゼC群およびその優性抑制変異体群の作用機構の解析

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] Hirai,S.,Izumi,Y.,Higa,K.,Mizuno,K.,Osada,S.,Kaibuchi,K.,Suzuki,K.and Ohno,S.: "Ras-dependent signal transducition is indispensable but not sufficient for the activation of AP1/Jun by PKCdelta." EMBOJ.(in press).

[文献書誌] Ohno,S.and Suzuki,K.: "Protein kinase C:Reassessment.Function of the novel members of the protein kinase C family." FESEB J.(in press).

[文献書誌] Chida,K.,Sagara,H.,Suzuki,Y.,Murakami,A.,Osada,S.,Ohno,S.,Hirosawa,K,and Kuroki,T.: "The eta isoform of protein kinase C(nPKCeta)is localized on rough endoplasmic reticulum." Molecular and Cellular Biol.(in press).

[文献書誌] Koyama,H.,Inaba,M.,Nishizawa,Y.,Ohno,S.and Morii,H.: "Protein kinase C is involved in 24-hydroxylase gene expression induced by 1,25(OH)_2D_3 in rat intestinal epithelial cells." J Cellular Biochemistry. (in press).

[文献書誌] Suzuki,K.,Akita,Y.and Ohno,S.: "Structural and functional diversities of protein kinase C family members." Recent Advances in Molecular and Biochemical Research on proteins.(in press).

[文献書誌] Ohno,S.,Akita,Y.,Hata,A.,Osada,S.,Kubo,K.,Konno,Y.,Akimoto,K.,Mizuno,K.,Saido,T.C.,Kuroki,T.and Suzuki,K.: "Structural and functional diversities of a family of signal transducing protein Kinases;protein Kinase C family." Yckohama Med.Bull.43. 139-140 (1993)

[文献書誌] Ohno,S.: "Diversity and specificity of multiple protein kinase C." Progress in Endocrinology,Parthenon Press. 63-66 (1993)

[文献書誌] Koizumi,H.,Kohno,Y.,Osada,S.,Ohno,s.,Ohkawara,and Kuroki,T.: "Differentiation-associated localization of nPKCeta,a Ca^<++>-independent protein kinase C,in normal human skin and skin diseases." J.Investigative Dermatology. 6. 858-863 (1993)

[文献書誌] Mischak,H.,Goodnight,J.,Henderson,D.W.,Osada,S.,Ohno,s.and Mushinski,F.: "Unique expression pattern of protein kinase C-theta:high _mRNA levels in normal mouse testes and in T-lymphocytic cells and neoplasms." FEBS Lett.326. 51-55 (1993)

[文献書誌] Sublette,E.,Naik,M.U.,Jiang,X,Osten,P.,Valsarmis,H.,Osada,S.,Ohno,S.and Sacktor,T.C.: "Evidence for a new,high-molecular weight isoform of protein kinase C in rat hippocampus." Neuroscience Lett.159. 175-178 (1993)

[文献書誌] Mizuno,K.,Saido,T.C.,Ohno,S.,Tamaoki,T.and Suzuki,K.: "Staurosporine-related compounds,K252a and UCN-01,inhibits both cPKC and nPKC." FEBS Lett.330. 114-116 (1993)

[文献書誌] Park,H.-Y.,Russakovsky,V.,Ohno,S.and Gilchrest,B.A.: "The beta-Isoform of protein kinase C stimulates human melanogenesis by act:rating tyrosinase in pigment cells." J.Biol.Chem.268. 11742-11749 (1993)

[文献書誌] 大野茂男: "Cキナーゼファミリーの活性調節" 細胞増殖の制御,南江堂. 105-114 (1993)

[文献書誌] 大野茂男: "細胞増殖と疾患-細胞増殖の細胞内シグナル伝達機構" Mebio,メジカルビュー社. 10,6. 30-38 (1993)

1994 年度実績報告書

大野茂男横浜市立大学, 医学部, 教授 (10142027)