对RNA结构的全基因组探测揭示了mRNA结构的活跃展开 in vivo
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
概括
在体内RNA结构监测显示,细胞内部的结构化mRNA区域比体内更少. 细胞过程,不仅仅是热力学,决定活体内的RNA折叠.
科学领域:
- 分子生物学分子生物学
- 遗传学 是一个遗传学.
- 生物化学 生物化学
背景情况:
- RNA分子执行各种生物功能,需要复杂的结构.
- 目前用于RNA结构确定的方法主要是体外检测,限制了体内应用.
- 在体内RNA折叠受蛋白质和酶等细胞组件的影响,而不仅仅是热力学.
研究的目的:
- 开发和应用一种新的方法,以单核酸分辨率在原生细胞条件下全球监测RNA结构.
- 为了比较体内和体外RNA结构,并研究影响细胞内RNA折叠的因素.
主要方法:
- 开发一种 in vivo RNA 结构探测技术,使用二甲基硫酸盐 (DMS) 修饰未配对基的方法.
- 深度测序DMS修改RNA以全基因组映射结构元素.
- 将体内RNA结构与已知的体外和晶体结构进行比较,并在缺乏ATP的条件下进行分析.
主要成果:
- 开发的方法准确地反映了酵母和哺乳动物细胞中已知的RNA结构.
- 与体外相比,体内观察到结构化mRNA区域的数量显著减少,即使对于热稳定结构.
- 发现依赖ATP的细胞过程在维持mRNA在体内主要展开的状态方面至关重要.
结论:
- 细胞环境积极调节RNA结构,偏离了简单的热力学预测.
- 在体内RNA结构是动态的,并受到能源依赖的细胞机械的影响.
- 这项研究提供了一个强大的工具,用于分析功能性RNA结构在它们的原生细胞环境中.
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