RNA構造の全ゲノム調査は,mRNA構造の活性展開を in vivo で明らかにしています
Silvi Rouskin1, Meghan Zubradt1, Stefan Washietl2
1Department of Cellular and Molecular Pharmacology, California Institute of Quantitative Biology, Center for RNA Systems Biology, Howard Hughes Medical Institute, University of California, San Francisco, California 94158, USA.
Nature
|December 17, 2013
まとめ
In vivo RNA構造モニタリングでは,細胞内の構造化されたmRNA領域がin vitroよりも少ないことが明らかになる. 熱力学だけでなく,細胞のプロセスも,生命体内のRNAの折り畳みを決定する.
科学分野:
- 分子生物学は分子生物学である.
- 遺伝学 遺伝学とは
- バイオケミストリー バイオケミストリー
背景:
- RNA分子は多様な生物学的機能を果たし,複雑な構造を必要とします.
- RNA構造の決定のための現在の方法は,主に in vitro で,in vivo の適用性を制限しています.
- In vivo RNAの折りたたみには,熱力学だけでなく,タンパク質やヘリコースなどの細胞成分が影響する.
研究 の 目的:
- シングル・ヌクレオチド解像度で,ネイティブの細胞環境におけるRNA構造をグローバルにモニタリングするための新しい方法を開発し,適用する.
- in vivoとin vitroのRNA構造を比較し,細胞内のRNA折り畳みに影響する要因を調査する.
主な方法:
- 非ペア化塩基のダイメチル硫酸 (DMS) 改変を用いたインビヴォRNA構造探査技術の開発.
- 構造要素をゲノム全体にマッピングするために,DMS改変RNAの深層配列化.
- in vivoのRNA構造と既知のin vitroおよび結晶構造を比較し,ATPが不足した条件下で分析した.
主要な成果:
- 開発された方法は,酵母と哺乳類の細胞の既知のRNA構造を正確に反映しています.
- 熱安定構造であっても,体内と比較して,体内では構造化されたmRNA領域が著しく少ないことが観察されました.
- ATPに依存する細胞プロセスは,mRNAの主に展開された状態を維持する上で,in vivoにおいて極めて重要であることが判明しました.
結論:
- 細胞環境はRNAの構造を積極的に調節し,単純な熱力学的予測から逸脱する.
- In vivo RNAの構造はダイナミックで,エネルギーに依存する細胞機構の影響を受けます.
- この研究は,本来の細胞文脈における機能的なRNA構造を分析するための強力なツールを提供します.
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