1993 年度実績報告書

細胞増殖におけるプロテインキナーゼC群の機能の分子生物学的解析

研究課題

研究課題/領域番号	04253239
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	秋田朗子東京都臨床医学総合研究所, 遺伝情報, 研究員 (40124432) 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 / ジアシルグリセロール
研究概要	昨年度来の研究から、我々は線維芽細胞への各PKC分子種の高発現系と転写応答のレポーター遺伝子とを用いて、生きた細胞内でのPKCの活性化状況をモニターする系を始めて開発した。この系を用いて本年度は新たに_aPKCの活性化をもモニター出来る事を確認した。この系を用いて、_cPKC、_nPKC、_aPKCという3種のPKC亜群の各々の分子種について、その活性化機構を解析することが始めて可能となった。そこで本年度はこの点を集中的に解析し、PKCの生理的な重要性の発見以来約10年ぶりともいえる画期的な発見をすることが出来た。それは一言でいうとPKCの活性化経路には、従来から知られていたPLC/DAGを介する経路以外に、PI3キナーゼを介する全く新しい経路が関わっているという点である。チロシンキナーゼ型受容体の代表格と言えるPDGF受容体のシグナル異変体や、PI3キナーゼサブユニットの発現系を用いて、各PKC分子種の活性化に関わる経路を生きた細胞を用いて分子生物学的に始めて解析し、従来のPLCを介した経路が_cPKC、_nPKCの活性化に関わることを確認すると同時に、PI3キナーゼを介する経路がこれらの活性化に関わることを始めて見いだした。更に、_aPKCの活性化にはPLCではなく、PI3キナーゼが関わることを始めて見いだした。

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 transduction 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,PKC lambda,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 aCa_+-independent protein kinase C isozyme,_nPKC epsilon,increases the secretion of protein from thyrotropin-releasing hormone-stimulated rat pituitary GII_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(_nPKC_<eta>)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." Yokohama 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,A.and Kuroki,T.: "Differentiation-associated localization of _nPKC eta,_aC_a^<++>-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.,Valsamis,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,K252_a 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 activating tyrosinase in pigment cells.21GC14: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:PKC_<gamma> differs from PKC alpha,betaII,and _nPKC epsilon 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.,Konno,Y.,Chida,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,_aC_a^<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 Kamocki,E.: "A new protein kinase C,_nPKC eta′and PKC theta are expressed in human platelets:involvement of _nPKC_n′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)

1993 年度 実績報告書

細胞増殖におけるプロテインキナーゼC群の機能の分子生物学的解析

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 transduction 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(_nPKC_<eta>)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." Yokohama 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,A.and Kuroki,T.: "Differentiation-associated localization of _nPKC eta,_aC_a^<++>-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.,Valsamis,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,K252_a 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 activating tyrosinase in pigment cells.21GC14: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)

1993 年度実績報告書

大野茂男横浜市立大学, 医学部, 教授 (10142027)