Envelope stress is a trigger of CRISPR RNA-mediated DNA silencing in Escherichia coli.
Envelope stress is a trigger of CRISPR RNA-mediated DNA silencing in Escherichia coli.
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DOI：
10.1111/j.1365-2958.2010.07482.x
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发表时间：
2011-02
影响因子：
3.6
通讯作者：
DeLisa MP
中科院分区：
文献类型：
--
作者：
Perez-Rodriguez R;Haitjema C;Huang Q;Nam KH;Bernardis S;Ke A;DeLisa MP
大多数细菌和古细菌基因组中的一个普遍特征是一系列成簇的、规则间隔的短回文重复序列（CRISPR），它们与一组CRISPR相关（Cas）蛋白一起介导针对侵入性核酸（如质粒和病毒）的免疫力。在此，CRISPR-Cas系统在表达靶向双精氨酸易位（达特）途径的质粒编码蛋白的细胞中被激活。这种达特底物的表达导致Cas酶的上调和随后的编码质粒的沉默，其方式需要BaeSR双组分调节系统，已知该系统响应于胞质外应激。此外，我们证实CasCDE酶形成稳定的三元复合物，并且似乎作为Cas系统的催化核心将CRISPR RNA加工成其成熟形式。总之，我们的结果表明，CRISPR-Cas系统直接靶向DNA，作为细菌防御机制的一部分，与噬菌体感染重叠但不限于噬菌体感染。
A widespread feature in the genomes of most bacteria and archaea is an array of clustered, regularly interspaced short palindromic repeats (CRISPRs) that, together with a group of CRISPR-associated (Cas) proteins, mediate immunity against invasive nucleic acids such as plasmids and viruses. Here, the CRISPR-Cas system was activated in cells expressing a plasmid-encoded protein that was targeted to the twin-arginine translocation (Tat) pathway. Expression of this Tat substrate resulted in upregulation of the Cas enzymes and subsequent silencing of the encoding plasmid in a manner that required the BaeSR two-component regulatotory system, which is known to respond to extracytoplasmic stress. Further, we confirm that the CasCDE enzymes form a stable ternary complex and appear to function as the catalytic core of the Cas system to process CRISPR RNA into its mature form. Taken together, our results indicate that the CRISPR-Cas system targets DNA directly as part of a defense mechanism in bacteria that is overlapping with but not limited to phage infection.
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