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/領域番号: 06283222
• 研究種目: 重点領域研究
• 配分区分: 補助金
• 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) ; 長田 真一 横浜市立大学, 医学部, 助手 (00244484) ; 平井 秀一 横浜市立大学, 医学部, 講師 (80228759)
• 研究期間 (年度): 1996
• 研究課題ステータス: 完了 (1996年度)
• 配分額: 37,000千円 (直接経費: 37,000千円); 1996年度: 15,000千円 (直接経費: 15,000千円); 1995年度: 11,000千円 (直接経費: 11,000千円); 1994年度: 11,000千円 (直接経費: 11,000千円)
• 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 / PI3キナーゼ / ホスホリパーゼC / プロテインキナーゼC結合タンパク質 / 血清飢餓 / DNA合成 / 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

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

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