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GAL4誘導体は単独で作用し,哺乳類の活性化剤と相乗効果で in vitro 作用する

Y S Lin¹, M F Carey, M Ptashne

¹Department of Biochemistry and Molecular Biology, Harvard University, Cambridge, Massachusetts 02138.

|August 26, 1988

まとめ

イーストのGAL4タンパク質誘導体は,哺乳類の遺伝子転写を in vitroで活性化させます. 効率的な活性化には,特定の結合部位の配置または哺乳類の活性化剤との相乗効果が必要であり,新しい in vitro 特定の活性化領域を明らかにします.

科学分野:

分子生物学は分子生物学である.
遺伝子規制遺伝子規制
プロテイン工学は,タンパク質の

背景:

イーストのトランスクリプションアクティベーターGAL4は,よく研究されたトランスクリプション因子です.
ユカリオット遺伝子の転写メカニズムを理解することは,様々な生物学的プロセスにとって極めて重要です.

研究の目的:

哺乳類のシステムにおけるGAL4誘導体のインビトロ転写活性化能力を調査する.
GAL4結合部位の位置と他の活性化剤との相互作用が転写刺激に及ぼす影響を決定する.

主な方法:

E. coliからのGAL4誘導体の合成と浄化.
HeLa細胞核抽出物と改変されたアデノウイルスE4遺伝子テンプレートを使用したインビトロ転写アッセイ.
E4 TATA ボックスに対する GAL4 結合部位の位置の体系的な変化.

主要な成果:

GAL4誘導体は,ヘラ核抽出物におけるアデノウイルスE4遺伝子転写を効率的に刺激した.
最適な刺激は,GAL4結合部位がTATAボックスのすぐ上流に配置されたときに発生した.
遠隔の GAL4 サイトでは,シナギスティックな刺激のために近隣の哺乳類のアクティベーターサイトを必要とし,文脈依存のアクティベーションを強調しました.

結論:

GAL4誘導体は,異種哺乳類のシステムで転写活性化剤として機能することができる.
この研究では,GAL4における第3の活性化領域が特定され,これはin vitroでのみ機能する.
発見は,転写活性化メカニズムのモジュール性と文脈依存性についての洞察を提供します.

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