p21によるPCNAの機能制御と細胞のがん化

• 研究課題/領域番号: 08265102
• 研究種目: 特定領域研究(A)
• 配分区分: 補助金
• 研究機関: 奈良先端科学技術大学院大学
• 研究代表者: 釣本 敏樹 奈良先端科学技術大学院大学, バイオサイエンス研究科, 助教授 (30163885)
• 研究分担者: 小布施 力史 奈良先端科学技術大学院大学, バイオサイエンス研究科, 助手 (00273855) ; 藤田 雅俊 愛知県がんセンター研究所, 研究員 (30270713) ; 大谷 清 東京医科歯科大, 疾患遺伝子実験センター, 助手 (30201974) ; 村上 洋太 京都大学, ウイルス研究所, 助教授 (20260622) ; 正井 久雄 東京大学, 医科学研究所, 助教授 (40229349) ; 加藤 順也 奈良先端科学技術大学院大学, バイオサイエンス研究科, 助教授 (00273839) ; 秋山 昌広 奈良先端科学技術大学院大学, バイオサイエンス研究科, 助教授 (80273837)
• 研究期間 (年度): 1996 – 1999
• 研究課題ステータス: 完了 (1999年度)
• 配分額: 110,400千円 (直接経費: 110,400千円); 1999年度: 7,500千円 (直接経費: 7,500千円); 1998年度: 37,000千円 (直接経費: 37,000千円); 1997年度: 33,000千円 (直接経費: 33,000千円); 1996年度: 32,900千円 (直接経費: 32,900千円)
• キーワード: DNA複製 / PCNA結合タンパク質 / 細胞周期 / S期 / DNAメチル転移酵素 / クロマチン集合 / DNA修復 / 染色体構造 / ORC / PCNA / 複製開始点結合因子 / S期導入因子 / ライセンシング因子 / MCM / 蛋白質リン酸化 / E2F-1

研究概要

PCNAはDNA複製の必須因子であると共に多様な因子と相互作用し、複製が進行した後のDNAの修復、染色体の再構築に深く関わることから、細胞の正常な増殖状態の維持に重要な役割を持つと考えられている。さらにこれにCDK/cyclinの阻害因子であるp21(Cip 1, Waf1)が結合し、その発現量に応じてPCNAの機能を制御する機構が想定されている。このPCNAの機能制御と正常な細胞増殖の状態の維持の関係を明らかにするため、PCNA結合タンパク質の解析およびこの結合に対するp21の影響を解析した。PCNA固定化カラムを用いてPCNA結合タンパク質を網羅的に検索した。その結果、既知を含め、10以上のPCNA結合タンパク質が検出された。この系にp21ペプチドを添加することで、p21によるPCNA結合タンパク質の結合制御を解析することができる。その結果、そのほとんどがp21により結合が抑制されること、その抑制の程度は、結合因子によって特異性があることが明らかになった。さらにPCNA結合因子の個々の特性について解析を行った。DNAメチル転移酵素MeTase)とPCNAの結合の特異性について解析し、PCNAのとの結合により(MeTase)のDNAへの相互作用が促進され、p21がこれを抑制することを示すことができた。MeTase、Fen1エンドヌクレース、クロマチン集合因子等についてPCNA上の結合部位を解析した。その結果、これらはp21やDNAポリメラーゼがPCNA上に結合するのと共通した部位に結合することを見い出し、p21の競合的結合によりこれらの結合が制御されることを明らかにした。以上より、S期核内でPCNAは複製の進行中の染色体構造維持に多面的機能を持ち、p21がそれに対する統括的制御の役割を持つことが示唆される。

研究成果

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