HIV-1 Revのペプチドによって,mRNAのスプライシングをインビトロで特定調節する
J Kjems1, A D Frankel, P A Sharp
1Center for Cancer Research, Massachusetts Institute of Technology, Cambridge 02139.
Cell
|October 4, 1991
まとめ
HIV-1のRevタンパク質は,Rev応答要素 (RRE) に結合することによって,ウイルスのmRNAのスプライシングを阻害する. Revを模倣する合成ペプチド.
科学分野:
- 分子生物学は分子生物学である.
- ウイルス学 ウイルス学 ウイルス学
- RNA 処理 RNA 処理
背景:
- ヒト免疫不全ウイルス1型 (HIV-1) のRevタンパク質は,ウイルスの遺伝子発現を調節するために重要である.
- Revは,核から細胞プラズマへの未結合および部分結合のウイルスのmRNAの輸出を容易にする.
- この調節は,RevがRev応答要素 (RRE) と呼ばれる特定のRNA構造との相互作用によって媒介されます.
研究 の 目的:
- Revタンパク質がpre-mRNA splicingをin vitroで調節するメカニズムを調査する.
- スプライシング阻害におけるRev RNA結合ドメインの役割を特徴づける.
- Rev-mediated splicingの構造要件を決定する.
主な方法:
- Rev応答要素 (RRE) を含むプレ-mRNAを用いたインビトロスプライシングアッセイ.
- Rev.のRNA結合ドメインを模倣する合成ペプチドの特徴化.
- 野生型のRevと,異なるRRE結合特性を有する合成ペプチドによるスプライシング阻害の分析.
主要な成果:
- Revタンパク質は,特に,RREを含むmRNA前スプライシングを3〜4倍抑制する.
- RevRNA結合ドメインを含む合成17アミノ酸ペプチドは,スプライシングを最大30倍抑制する.
- ペプチドの複数の結合部位は,RREの代用となり,多価相互作用の重要性を示している.
- ペプチドの抑制作用は,スプライシング複合体の組み立て中に発生し,スプライソーム形成への干渉を示唆しています.
結論:
- RevのRNA結合ドメインは,スペライシングの特定の阻害に十分である.
- Rev-mediated splicing阻害は,RRE上の複数の結合部位との相互作用を伴う.
- Revの基本領域は,機能的なスプライソームの形成を防ぐために機能し,それによってウイルスのmRNA処理を調節します.
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