1996 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	鈴木厚横浜市立大学, 医学部, 助手 (00264606) 水野恵子横浜市立大学, 医学部, 助手 (90221803)
キーワード	プロテインキナーゼC / S6キナーゼ / ストレス応答 / NFkB / メタロプロティナーゼ
研究概要	PKCは細胞内シグナル経路の調節因子として極めて重要であることが様々な実験から推測されてきたが、未だにその具体的な作用点は不明である。本研究では、従来の研究で集積したPKCに関する独自の材料を用いて、PKCの具体的な作用点を解明することを目的とし、以下の二つの方向からのアプローチを行い、以下の結果を得た。 1)PI3キナーゼ経路の代表的な下流因子、p70S6キナーゼの活性化機構を解析し、これにaPKCλが関わっていることを明らかにした。 2)nPKCδ結合蛋白質として細胞膜貫通型のタンパク質を同定した。これはメタロプロテイナーゼ/ディスインテグリンファミリーの仲間であり、PKCδはこの細胞質ドメインに特異的に結合する。細胞接着とシグナル伝達経路との接点となっている事が予測され、きわめて興味深い。 3)aPKCλ結合タンパク質として、PDZドメインを有する新規タンパク質を見いだした。これは線虫の卵の不等分裂に関わる遺伝子のほ乳類版であった。 4)Rafを介したMAPキナーゼの活性化過程において、Ras以外に必要な因子として、PKCδを同定した。 5)ストレス応答MAPキナーゼ(SAPK/JNK)経路の上流キナーゼとして、MUK、MST/MLK2を同定し、これらがSEKを介してSAPK/JNKを活性化することを示した。SAPK/JNKの上流には、MEKKに加えてMUK/MLKファミリーが存在する。 6)SAPK/JNK上流因子の一つであるMEKKがTNFαによるNFkBの活性化経路に関わることを見出した。

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] Fukumoto, S.: "Protein kinase C delta isoform inhibits the proliferation of vascular smooth muscle cells through suppressiong G1 cyclins expression." J.Biol.Chem.272(in press.).
[文献書誌] Hirai, S.: "MST/MLK2, a member of the mixed lineage kinase family, directly phosphorylates and activates SEK1, an activator of JNK/SAPK." J.Biol.Chem.272(in press.).
[文献書誌] Izumi, Y.: "A protein kinase C delta-binding protein SRBC whose expression is induced by serum starvation." J.Biol.Chem.272(in press.).
[文献書誌] Osada, S.: "A domain containing the Cdc42/Rac interactive binding (CRIB) region of p65PAK inhibits transcriptional activation and cell transformation mediated by the Ras-Rac pathway." FEBS Letter. (in press.).
[文献書誌] Osada, S.: "YSK1, a novel mammalian protein kinase structurally related to Ste20 and SPS1, but not involved in the known MAPK pathways." Oncogene. 14(in press.).
[文献書誌] Ohno, S.: "The distinct biological potential of PKC isotypes." in Protein Kinase C. 75-93 (1996)
[文献書誌] Zappelli, F.: "The inhibition of differentiation caused by TGFbeta in fetal myoblasts is dependent upon selective expression of PKCtheta : a possible molecular basis for myoblast diversification during limb histogenesis." Dev.Biol.180. 156-164 (1996)
[文献書誌] Takayanagi, I.: "Dominant negative effects of the trucated p110 subunit of phosphatidylinositol-3 kinase." Biochem. Mol.Biol.Int.39. 721-728 (1996)
[文献書誌] Chalfant, C.E.: "A carboxy-terminal deletion mutant of PKCbeta II inhibits insulin-stimulated 2-deoxyglucose uptake in rat L6 skeletal muscle cells." Molecular Endocrinology. 10. 1273-1281 (1996)
[文献書誌] Chalfant, C.E.: "A carboxy-terminal deletion mutant of PKCbeta II inhibits insulin-stimulated 2-deoxyglucose uptake in rat L6 skeletal muscle cells." Molecular Endocrinology. (in press.).
[文献書誌] Ueda, Y.: "Protein kinase Cδ activates the MEK-ERK pathway in a manner independent of Ras and dependent on Raf." J.Biol.Chem.271. 23512-23519 (1996)
[文献書誌] Rana, A.: "The mixed lineage kinase SPRK phosphorylates and activates the stress-activated protein kinase activator, SEK-1." J.Biol.Chem.271. 19025-19028 (1996)
[文献書誌] Sauma. S.: "Protein kinase Cβ1 and protein kinase Cβ2 activate p57 mitogen-activated protein kinase and block differentiation in colon carcinoma cells." Cell Growth Differentiation. 7. 587-594 (1996)
[文献書誌] Nakaigawa, N.: "Differential effects of overexpression of PKCα and PKCδ/ε on cellular E2F activity at late G1 phase." Biochem. Biophys. Res.Commun.222. 95-100 (1996)
[文献書誌] Hirano, M.: "MEK kinase is involved in TNFα-mediated activation of NFκB and degradation of IκBα." J.Biol.Chem.271. 13234-13238 (1996)
[文献書誌] Ueda, E.: "The η isoform of protein kinase C mediates transcriptional activation of the human transglutaminase 1 gene." J.Biol.Chem.271. 9790-9794 (1996)
[文献書誌] Akimoto, K.: "EGF and PDGF receptors activate atypical PKCλ through phosphatidylinositol 3-kinase." EMBO J.15. 788-798 (1996)
[文献書誌] Hirai, S.: "Activation of the JNK pathway by distantly related protein kinases, MEKK and MUK." Oncogene. 12. 641-650 (1996)
[文献書誌] Moriya, S.: "PDGF activates protein kinase Cε through redundant and independent signaling pathways involving phosphpholipase Cγ or phosphatidylinositol 3-kinase." Proc Natl.Acad.Sci.U.S.A.93. 151-155 (1996)

1996 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] Fukumoto, S.: "Protein kinase C delta isoform inhibits the proliferation of vascular smooth muscle cells through suppressiong G1 cyclins expression." J.Biol.Chem.272(in press.).

[文献書誌] Hirai, S.: "MST/MLK2, a member of the mixed lineage kinase family, directly phosphorylates and activates SEK1, an activator of JNK/SAPK." J.Biol.Chem.272(in press.).

[文献書誌] Izumi, Y.: "A protein kinase C delta-binding protein SRBC whose expression is induced by serum starvation." J.Biol.Chem.272(in press.).

[文献書誌] Osada, S.: "A domain containing the Cdc42/Rac interactive binding (CRIB) region of p65PAK inhibits transcriptional activation and cell transformation mediated by the Ras-Rac pathway." FEBS Letter. (in press.).

[文献書誌] Osada, S.: "YSK1, a novel mammalian protein kinase structurally related to Ste20 and SPS1, but not involved in the known MAPK pathways." Oncogene. 14(in press.).

[文献書誌] Ohno, S.: "The distinct biological potential of PKC isotypes." in Protein Kinase C. 75-93 (1996)

[文献書誌] Zappelli, F.: "The inhibition of differentiation caused by TGFbeta in fetal myoblasts is dependent upon selective expression of PKCtheta : a possible molecular basis for myoblast diversification during limb histogenesis." Dev.Biol.180. 156-164 (1996)

[文献書誌] Takayanagi, I.: "Dominant negative effects of the trucated p110 subunit of phosphatidylinositol-3 kinase." Biochem. Mol.Biol.Int.39. 721-728 (1996)

[文献書誌] Chalfant, C.E.: "A carboxy-terminal deletion mutant of PKCbeta II inhibits insulin-stimulated 2-deoxyglucose uptake in rat L6 skeletal muscle cells." Molecular Endocrinology. 10. 1273-1281 (1996)

[文献書誌] Chalfant, C.E.: "A carboxy-terminal deletion mutant of PKCbeta II inhibits insulin-stimulated 2-deoxyglucose uptake in rat L6 skeletal muscle cells." Molecular Endocrinology. (in press.).

[文献書誌] Ueda, Y.: "Protein kinase Cδ activates the MEK-ERK pathway in a manner independent of Ras and dependent on Raf." J.Biol.Chem.271. 23512-23519 (1996)

[文献書誌] Rana, A.: "The mixed lineage kinase SPRK phosphorylates and activates the stress-activated protein kinase activator, SEK-1." J.Biol.Chem.271. 19025-19028 (1996)

[文献書誌] Sauma. S.: "Protein kinase Cβ1 and protein kinase Cβ2 activate p57 mitogen-activated protein kinase and block differentiation in colon carcinoma cells." Cell Growth Differentiation. 7. 587-594 (1996)

[文献書誌] Nakaigawa, N.: "Differential effects of overexpression of PKCα and PKCδ/ε on cellular E2F activity at late G1 phase." Biochem. Biophys. Res.Commun.222. 95-100 (1996)

[文献書誌] Hirano, M.: "MEK kinase is involved in TNFα-mediated activation of NFκB and degradation of IκBα." J.Biol.Chem.271. 13234-13238 (1996)

[文献書誌] Ueda, E.: "The η isoform of protein kinase C mediates transcriptional activation of the human transglutaminase 1 gene." J.Biol.Chem.271. 9790-9794 (1996)

[文献書誌] Akimoto, K.: "EGF and PDGF receptors activate atypical PKCλ through phosphatidylinositol 3-kinase." EMBO J.15. 788-798 (1996)

[文献書誌] Hirai, S.: "Activation of the JNK pathway by distantly related protein kinases, MEKK and MUK." Oncogene. 12. 641-650 (1996)

[文献書誌] Moriya, S.: "PDGF activates protein kinase Cε through redundant and independent signaling pathways involving phosphpholipase Cγ or phosphatidylinositol 3-kinase." Proc Natl.Acad.Sci.U.S.A.93. 151-155 (1996)

1996 年度実績報告書

大野茂男横浜市立大学, 医学部, 教授 (10142027)