in vivoの構造的インプリントは,RNAの規制メカニズムをデコードする
Robert C Spitale1, Ryan A Flynn1, Qiangfeng Cliff Zhang1
1Howard Hughes Medical Institute and Program in Epithelial Biology, Stanford University School of Medicine, Stanford, California 94305, USA.
Nature
|March 25, 2015
まとめ
研究者は,生きている細胞のRNA構造をマッピングするために,選択的2'-ヒドロキシルアシレーションとプロファイリング実験 (icSHAPE) をin vivoで開発しました. この方法は,遺伝子発現とRNAとタンパク質の相互作用に不可欠なダイナミックなRNA構造を明らかにします.
科学分野:
- 分子生物学は分子生物学である.
- ゲノミクスゲノミクスとは
- バイオケミストリー バイオケミストリー
背景:
- 生体細胞内の分子行動を視覚化することは,重要な生物学的課題です.
- RNA構造は遺伝子発現の調節に不可欠ですが,現在のin vivo方法では不完全なプロファイルが提供されています.
- 生理学的RNA構造の理解は限られており,既存の技術は4つのRNA塩基のうち2つしかカバーしていない.
研究 の 目的:
- 生体細胞におけるグローバルRNA二次構造プロファイリングのための新しい生化学的アプローチを提示する.
- 細胞環境内の4つのRNA塩基の構造の最初の包括的な見方を可能にするために.
- in vivoとin vitroのRNA構造ダイナミクスの違いを調査する.
主な方法:
- in vivo クリック選択型2 -ヒドロキシルアシレーションとプロファイリング実験 (icSHAPE) の開発と応用.
- マウスの胚性幹細胞におけるグローバルトランスクリプトーム幅のRNA構造のプロファイリング.
- in vivoとin vitroで得られたRNA構造の比較.
主要な成果:
- icSHAPEは,生きている細胞の4つの塩基すべてのためのRNA二次構造の最初のグローバルビューを提供します.
- In vivo RNAの構造的動態は,in vitroの条件とは異なり,RNAのクラスと規制要素を区別する.
- 翻訳部とリボソームの休止部位の構造的シグネチャーが保存され,シーケンスプログラミングを示す.
- ダイナミックな構造的足跡は,RNA-タンパク質相互作用インターフェースとRNA改変部位を明らかにし,原子データと一致しています.
- RNA-タンパク質相互作用とN(6) -メチラデノシン (m(6) A) 改変の正確なゲノム全体の予測が可能になります.
結論:
- icSHAPE技術は,生きている細胞におけるRNA構造ゲノミクスの新しい道を開きます.
- 生理学的RNA構造は,遺伝子発現を制御する上で重要な役割を果たします.
- この研究は,タンパク質と変化の相互作用を媒介する正確なRNA構造動態を明らかにしています.
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