DNA合成酵素遺伝子の構造と機能

1989 年度 実績報告書

• 研究課題/領域番号: 01010084
• 研究機関: 愛知県がんセンター
• 研究代表者: 松影 昭夫 愛知県がんセンター研究所, 生物学部, 部長 (90019571)
• 研究分担者: 有賀 寛芳 北海道大学, 薬学部・微生物薬品化学, 教授 (20143505) ; 安藤 俊夫 愛知県がンセンター研究所, 生化学部, 部長 (20012693) ; 竹石 桂一 静岡県立大学, 食品栄養科学部, 教授 (90012608) ; 小祝 修 愛知県コロニー発達障害研究所, 生化学部, 主任研究員 (50132923) ; 花岡 文雄 理化学研究所, 放射線生物学部, 主任研究員 (50012670)
• キーワード: DNA合成酵素の遺伝子 / DNAポリメラーゼ / 遺伝子発現調節 / 発現調節領域 / サイレンサー / 細胞増殖と分化 / c-Fosタンパク質 / C-Mycタンパク質

研究概要

DNA合成酵素遺伝子の機能的な解析が進展し、以下に示す実績が新たに得られた。(1)DNAポリメラーゼβ遺伝子のサイレンサーに結合する因子にがん遺伝子cーfosの産物が含まれることおよびこの結合領域の塩基配列はcーmyc遺伝子のサイレンサーと相同性をもつ。(2)ショウジョウバエのDNAポリメラーゼα遺伝子をクローニングして、その全配列をほぼ決定した。また、この遺伝子の上流にはPCNA遺伝子と同様のホメオドメインタンパク質の結合配列が存在することを見いだした。(3)DNAポリメラーゼαープライマーゼ複合体を構成するすでにクローン化された中心ポリペプチド以外の4つのポリペプチドの部分アミノ酸配列を決定し、cDNAと遺伝子のクローニングを進めている。(4)ターミナルトランスフェラーゼ遺伝子の発現調節領域にあるパリンドローム配列に特異的に結合する因子を見いだした。(5)ヒト先天性奇形において、PCNA、DNAポリメラーゼαとβ遺伝子に構造異常のある例をさらに見いだし、形態形成におけるDNA合成酵素の機能に示唆を与えた。(6)B型肝炎ウイルス逆転写酵素を支配する3.6kbmRNAのプロモーター結合タンパク質が肝細胞に特異的な因子と他の細胞にもある因子の複合体であることを証明した。(7)野性型およびカンプトテシン耐性のヒトトポイソメラーゼIcDNAの塩基配列を比較し、耐性をもたらしたのが、533番のGlyのAspへの変化であることを証明した。また、トポイソメラーゼIの活性変化によって核内のシグナル伝達系の変調がもたらされることを示唆した。(8)cーMycタンパク質が結合するcーmyc遺伝子上流にある配列とエンハンサーおよびDNA複製開始領域の関係をさらに詳細に解析した。また、cーMycのDNAへの結合にDNAポリメラーゼαも関与していることを見いだした。

研究成果

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