Viral recombination systems limit CRISPR-Cas targeting through the generation of escape mutations.
Viral recombination systems limit CRISPR-Cas targeting through the generation of escape mutations.
复制标题
病毒重组系统通过产生逃逸突变来限制CRISPR-Cas靶向。
DOI：
10.1016/j.chom.2021.09.001
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发表时间：
2021-10-13
影响因子：
30.3
通讯作者：
Marraffini LA
中科院分区：
文献类型：
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作者：
Hossain AA;McGinn J;Meeske AJ;Modell JW;Marraffini LA
CRISPR-Cas系统通过用识别和切割感染病毒基因组的RNA向导编程Cas核酸酶来提供对细菌的免疫力。细菌和它们的病毒各自编码能够修复被切割的病毒DNA的重组系统。然而，目前尚不清楚这些系统是否以及如何影响CRISPR免疫力。噬菌体λ使用Red系统（gam-exo-bet）来促进相关噬菌体之间的重组。在这里，我们表明λ Red也介导CRISPR-Cas靶向的逃避。Gam抑制宿主E. coli RecBCD重组系统，允许通过噬菌体Exo-Beta重组和修复切割的DNA，这促进了CRISPR靶序列内突变的产生。Red重组在产生大量逃避CRISPR靶向的RecBCD-RecA方面明显比宿主的RecBCD-RecA更有效。这些结果揭示了Red样系统在保护噬菌体对抗序列特异性核酸酶中的作用，这可能有助于它们在病毒基因组中的传播。在RNA引导的Cas核酸酶的靶位点处携带突变的噬菌体可以逃避CRISPR免疫。Hossain等人报道，噬菌体λ Red重组系统抑制宿主细菌RecBCD途径，并引入更有效地介导逃逸的突变。这些结果表明，噬菌体重组系统抵消DNA裂解细菌防御。
CRISPR-Cas systems provide immunity to bacteria by programming Cas nucleases with RNA guides that recognize and cleave infecting viral genomes. Bacteria and their viruses each encode recombination systems that could repair the cleaved viral DNA. However, it is unknown whether and how these systems can affect CRISPR immunity. Bacteriophage λ uses the Red system (gam-exo-bet) to promote recombination between related phages. Here we show that λ Red also mediates evasion of CRISPR-Cas targeting. Gam inhibits the host E. coli RecBCD recombination system, allowing recombination and repair of the cleaved DNA by phage Exo-Beta, which promotes the generation of mutations within the CRISPR target sequence. Red recombination is strikingly more efficient than the host’s RecBCD-RecA in the production of large numbers of phages that escape CRISPR targeting. These results reveal a role for Red-like systems in the protection of bacteriophages against sequence-specific nucleases, which may facilitate their spread across viral genomes. Bacteriophages harboring mutations at the target site of RNA-guided Cas nucleases can escape CRISPR immunity. Hossain et al. report that phage λ Red recombination system inhibits the host bacteria RecBCD pathway and introduces mutations that more efficiently mediate escape. These results suggest that phage recombination systems counteract DNA-cleaving bacterial defenses.
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