在体内结构性印记解码RNA调节机制
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
概括
研究人员在体内开发了选择性2'-基化和分析实验 (icSHAPE),以绘制活细胞中的RNA结构. 这种方法揭示了动态RNA结构,对于基因表达和RNA-蛋白相互作用至关重要.
科学领域:
- 分子生物学分子生物学
- 基因组学就是基因组学.
- 生物化学 生物化学
背景情况:
- 想象活细胞内的分子行为是一个重要的生物学挑战.
- RNA结构对于基因表达调节至关重要,但当前的体内方法提供不完整的概况.
- 了解生理学RNA结构是有限的,现有的技术只涵盖了四个RNA基中的两个.
研究的目的:
- 提出一种新的生物化学方法,用于在活细胞中分析全球RNA二次结构.
- 为了使细胞环境中所有四个RNA基的结构的第一个全面的视图.
- 为了研究体内和体外RNA结构动态之间的差异.
主要方法:
- 开发和应用体内选择性2-基化和分析实验 (icSHAPE).
- 在小鼠胚胎干细胞中的全球转录组全方位RNA结构概况.
- 在体内与体外获得的RNA结构的比较.
主要成果:
- icSHAPE为活细胞中所有四个基的RNA二次结构提供了第一个全球视图.
- 在体内RNA的结构动态不同于体内条件,区分RNA类别和调节元素.
- 转化和核糖体暂停部位的结构特征被保存,表明了序列编程.
- 动态结构足迹揭示了RNA-蛋白相互作用接口和RNA修饰部位,与原子数据一致.
- 能够准确的基因组范围预测RNA-蛋白相互作用和N(6) -甲基氨酸 (m(6) A) 修饰.
结论:
- icSHAPE技术为活细胞中的RNA结构基因组学开辟了新的途径.
- 生理学RNA结构在控制基因表达方面发挥着关键作用.
- 这项研究揭示了精确的RNA结构动态,它调解了与蛋白质和修饰的相互作用.
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