结构生物学. 结构生物学. 一个预先组织的Cas9指导RNA复合体,用于目标DNA识别
Fuguo Jiang1, Kaihong Zhou2, Linlin Ma2
1Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720, USA.
概括
细菌CRISPR-Cas9系统使用导向RNA的种子序列来预先订购用于DNA向的Cas9酶. 这种机制对于细菌适应性免疫至关重要,类似于真核细胞RNA干扰通路.
科学领域:
- 分子生物学分子生物学
- 微生物学 微生物学
- 遗传学 是一个遗传学.
背景情况:
- 细菌适应性免疫依赖于CRISPR-Cas系统来降解外来DNA.
- 包括Cas9内核酶在内的II型CRISPR-Cas系统通过不清楚的机制通过指导RNA结合来激活DNA识别.
研究的目的:
- 为了阐明Cas9激活在指导RNA结合时的结构机制.
- 了解引导RNA的种子序列是如何影响Cas9构造和DNA参与的.
主要方法:
- 确定Cas9与单导向RNA结合的晶体结构.
- 分析了Cas9在apo,指导RNA结合和DNA结合状态中的构造.
主要成果:
- 与单导向RNA结合的Cas9采用了与Apo和DNA结合状态截然不同的独特构造.
- 十核酸导向RNA种子序列采用A形形态,预先订购用于DNA查询.
- 这种预先订购的种子序列对于形成具有DNA识别能力的Cas9结构至关重要.
结论:
- 导向RNA种子序列在Cas9初始化中起着关键作用,用于目标DNA识别.
- 细菌CRISPR-Cas9中的这种"种子"机制在RNA干扰中显示出与真核细胞阿尔戈诺特蛋白的融合进化.
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