对CRISPR-Csm复合物的结构研究揭示了共转录干扰的机制
Lilan You1, Jun Ma2, Jiuyu Wang3
1Key Laboratory of RNA Biology, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, 100101 Beijing, China; University of Chinese Academy of Sciences, 100049 Beijing, China.
Cell
|December 4, 2018
概括
在CRISPR-Cas Csm复合体中
科学领域:
- 分子生物学
- 结构生物学
- 生物化学
背景情况:
- 这是一个CRISPR-Cas干扰系统.
- 它的功能包括RNA引导的RNA裂变,DNA裂变和循环甲酸 (cOA) 合成.
- 这些功能的目标RNA依赖激活的结构基础是未知的.
研究的目的:
- 阐明由目标RNA激活Csm复合物的结构机制.
- 了解目标RNA结合如何触发DNA裂变和cOA生成.
主要方法:
- 用X射线结晶学来确定Csm复合物的结构.
- 用于结构分析的冷电子显微镜 (Cryo-EM).
- 生物化学测定以评估酶活性.
主要成果:
- 确定了Csm复合体与crRNA和标RNA (同源和非同源) 的结构.
- 目标RNA的3'抗标签区域与Csm复合体上的两个不同的位点结合.
- 通过Csm1形态变化诱导DNA裂变和cOA合成的异质激活.
结论:
- 对crRNA介导的RNA裂变,目标RNA依赖的DNA裂变和cOA生成的结构性见解.
- 通过相关向RNA阐明了Csm复合体的向激活机制.
- 为理解III-A型CRISPR-Cas系统提供了一个框架.
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