TGF-β-DPC-4による細胞周期制御機構の解析

• 研究課題/領域番号: 09470042
• 研究種目: 基盤研究(B)
• 配分区分: 補助金
• 応募区分: 一般
• 研究分野: 病態医化学
• 研究機関: 東京大学 (1998); 大阪大学 (1997)
• 研究代表者: 秋山 徹 東京大学, 分子細胞生物学研究所, 教授 (70150745)
• 研究分担者: 石館 宇夫 東京大学分子細胞生物学研究所, 助手
• 研究期間 (年度): 1997 – 1998
• 研究課題ステータス: 完了 (1998年度)
• 配分額: 13,200千円 (直接経費: 13,200千円); 1998年度: 4,900千円 (直接経費: 4,900千円); 1997年度: 8,300千円 (直接経費: 8,300千円)
• キーワード: DPC4 / TGF-β / 標的遺伝子 / one-hybrid / サブトラクション / 細胞周期 / DPC-4, / TGF-b, / TAK1, / TAB-1, / 増殖制御、 / 細胞周期、 / シグナル伝達、 / 転写制御

研究概要

DPC4(SMAD4)はSAMD2あるいはSMAD3と複合体を形成し、TGF-β標的遺伝子のプロモーター領域に直接結合することで、標的遺伝子の発現を制御する。そこで、DNAと直接相互作用するDPC4をbaitに用いたyeast one-hybrid法を行うことでDPC4の標的遺伝子の探索を行った(one-hybrid法はbaitタンパク質と特異的に相互作用するDNA断片を単離する方法)。約2×10^5種類のヒトゲノム由来DNA断片をスクリーニングした結果、3種類のDNA断片を得た。One-hybrid法で得られたDNA断片内にはDPC4が認識するコンセンサス配列が実際に含まれており、本方法が標的遺伝子の探索に有効な方法のひとつであることが確認された。塩基配列決定後のデータベース検索により、1つがヒト28SrRNA遺伝子と一致することがわかった。残る2つのDNA断片については、目下解析中である。DPC4標的遺伝子の探索の別のアプローチとして、サブトラクション法及びプロモーター・トラップ法によるスクリーニングも進めており、現在、細胞周期・シグナル伝達・細胞骨格関連の遺伝子群と新規遺伝子群に焦点を定めた解析を行っている。さらに、DPC4による細胞周期制御機構を明らかにする目的でDPC4に特異的に結合する新規蛋白質の探索も昨年度に引き続き行っている。

研究成果

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  • 説明: 「研究成果報告書概要(欧文)」より
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  • 説明: 「研究成果報告書概要(欧文)」より
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  • 説明: 「研究成果報告書概要(欧文)」より
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