A dual function of the CRISPR-Cas system in bacterial antivirus immunity and DNA repair.
A dual function of the CRISPR-Cas system in bacterial antivirus immunity and DNA repair.
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DOI：
10.1111/j.1365-2958.2010.07465.x
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发表时间：
2011-01
影响因子：
3.6
通讯作者：
Yakunin AF
中科院分区：
文献类型：
--
作者：
Babu M;Beloglazova N;Flick R;Graham C;Skarina T;Nocek B;Gagarinova A;Pogoutse O;Brown G;Binkowski A;Phanse S;Joachimiak A;Koonin EV;Savchenko A;Emili A;Greenblatt J;Edwards AM;Yakunin AF
定期间隔的短静脉体重复序列（CRISPRS）和相关的蛋白质（CAS）组成了一种针对病毒和质粒的适应性免疫学系统，这是CAS1的CAS1蛋白质。在这里，我们表明，纯化的CAS1蛋白（YGBT）表现出对单链和分支DNA的核酸酶活性，包括霍利迪交界处，复制叉和5'-flaps。潜在的活性位点，全基因组筛选表明，YGBT与DNA修复系统的关键组成部分相互作用，包括RECB，RECC和RUVB一致。在CRISPR簇缺失的菌株中观察到了类似的表型，这表明YGBT在维修中的功能涉及与CRISPR的相互作用。除了抗病毒免疫外，CRISPR-CAS系统的至少某些组成部分在DNA修复中具有功能。
Clustered Regularly Interspaced Short Palindromic Repeats (CRISPRs) and the associated proteins (Cas) comprise a system of adaptive immunity against viruses and plasmids in prokaryotes. Cas1 is a CRISPR-associated protein that is common to all CRISPR-containing prokaryotes but its function remains obscure. Here we show that the purified Cas1 protein of Escherichia coli (YgbT) exhibits nuclease activity against single-stranded and branched DNAs including Holliday junctions, replication forks, and 5′-flaps. The crystal structure of YgbT and site-directed mutagenesis have revealed the potential active site. Genome-wide screens show that YgbT physically and genetically interacts with key components of DNA repair systems, including recB, recC and ruvB. Consistent with these findings, the ygbT deletion strain showed increased sensitivity to DNA damage and impaired chromosomal segregation. Similar phenotypes were observed in strains with deletion of CRISPR clusters, suggesting that the function of YgbT in repair involves interaction with the CRISPRs. These results show that YgbT belongs to a novel, structurally distinct family of nucleases acting on branched DNAs and suggest that, in addition to antiviral immunity, at least some components of the CRISPR-Cas system have a function in DNA repair.
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影响因子：
48
作者：
Butland, Gareth;Babu, Mohan;Emili, Andrew
通讯作者：
Emili, Andrew
影响因子：
11.4
作者：
Abraham, J;Lemmers, B;Hakem, R
通讯作者：
Hakem, R
影响因子：
11.4
作者：
Constantinou, A;Chen, XB;West, SC
通讯作者：
West, SC
影响因子：
14.9
作者：
Holm, L;Sander, C
通讯作者：
Sander, C
DOI：
10.1073/pnas.70.4.1064
发表时间：
1973-01-01
影响因子：
11.1
作者：
COLE, RS
通讯作者：
COLE, RS