Anti-CRISPR protein AcrIF4 inhibits the type I-F CRISPR-Cas surveillance complex by blocking nuclease recruitment and DNA cleavage.
Anti-CRISPR protein AcrIF4 inhibits the type I-F CRISPR-Cas surveillance complex by blocking nuclease recruitment and DNA cleavage.
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DOI：
10.1016/j.jbc.2022.102575
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发表时间：
2022-11
影响因子：
4.8
通讯作者：
Feng, Yue
中科院分区：
文献类型：
--
作者：
Gao, Zhengyu;Zhang, Laixing;Ge, Zihao;Wang, Hao;Yue, Yourun;Jiang, Zhuobing;Wang, Xin;Xu, Chenying;Zhang, Yi;Yang, Maojun;Feng, Yue
成簇的规则间隔短回文重复序列（CRISPR）-Cas系统为原核生物提供了针对移动的遗传元件（如peptide）的保护。反过来，CRISPR会部署抗CRISPR（Acr）蛋白来逃避这种免疫力。AcrIF 4是一种靶向I-F型CRISPR-Cas系统的Acr，据报道可结合crRNA引导的监视（Csy）复合物。然而，AcrIF 4是否抑制靶DNA与Csy复合物的结合仍存在争议。在这里，我们介绍了对AcrIF 4的结构和机制研究，探索其独特的抗CRISPR机制。虽然Csy-AcrIF 4复合物显示出对靶DNA的亲和力降低，但它仍然能够结合DNA。我们对Csy-AcrIF 4-dsDNA复合物的结构和功能分析表明，AcrIF 4结合阻止了由dsDNA结合诱导的Cas 8 f亚基的螺旋束的旋转，因此导致核酸酶Cas 2/3募集和DNA切割的失败。总的来说，我们的研究提供了一个有趣的例子，攻击核酸酶募集事件的Acr，但不是传统的机制，阻止结合的靶DNA。
The clustered regularly interspaced short palindromic repeats (CRISPR)-Cas system provides prokaryotes with protection against mobile genetic elements such as phages. In turn, phages deploy anti-CRISPR (Acr) proteins to evade this immunity. AcrIF4, an Acr targeting the type I-F CRISPR-Cas system, has been reported to bind the crRNA-guided surveillance (Csy) complex. However, it remains controversial whether AcrIF4 inhibits target DNA binding to the Csy complex. Here, we present structural and mechanistic studies into AcrIF4, exploring its unique anti-CRISPR mechanism. While the Csy–AcrIF4 complex displays decreased affinity for target DNA, it is still able to bind the DNA. Our structural and functional analyses of the Csy–AcrIF4–dsDNA complex revealed that AcrIF4 binding prevents rotation of the helical bundle of the Cas8f subunit induced by dsDNA binding, therefore resulting in failure of nuclease Cas2/3 recruitment and DNA cleavage. Overall, our study provides an interesting example of attack on the nuclease recruitment event by an Acr, but not conventional mechanisms of blocking binding of target DNA.
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影响因子：
5.9
作者：
Sanchez-Garcia R;Gomez-Blanco J;Cuervo A;Carazo JM;Sorzano COS;Vargas J
通讯作者：
Vargas J
影响因子：
2.5
作者：
Schindelin J;Rueden CT;Hiner MC;Eliceiri KW
通讯作者：
Eliceiri KW
影响因子：
4.8
作者：
Ren, Junhui;Wang, Hao;Yang, Lingguang;Li, Feixue;Wu, Yao;Luo, Zhipu;Chen, Zeliang;Zhang, Yi;Feng, Yue
通讯作者：
Feng, Yue
影响因子：
64.8
作者：
Bondy-Denomy J;Garcia B;Strum S;Du M;Rollins MF;Hidalgo-Reyes Y;Wiedenheft B;Maxwell KL;Davidson AR
通讯作者：
Davidson AR
DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
Emsley, P;Cowtan, K
通讯作者：
Cowtan, K