A bacteriophage nucleus-like compartment shields DNA from CRISPR nucleases
A bacteriophage nucleus-like compartment shields DNA from CRISPR nucleases
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DOI：
10.1038/s41586-019-1786-y
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发表时间：
2020-01-09
期刊：
影响因子：
64.8
通讯作者：
Bondy-Denomy, Joseph
中科院分区：
文献类型：
--
作者：
Mendoza, Senen D.;Nieweglowska, Eliza S.;Bondy-Denomy, Joseph
所有病毒都需要策略来抑制或逃避它们感染的细胞的免疫途径。感染细菌的病毒、噬菌体（噬菌体）必须避开靶向核酸的免疫途径，例如CRISPR-Cas和限制修饰系统，才能有效复制（1）。在这里，我们表明，巨型噬菌体phi KZ分离其宿主，铜绿假单胞菌的免疫核酸酶的DNA，通过构建一个蛋白质的核样隔室。phi KZ对体内靶向DNA的许多免疫机制具有抗性，包括CRISPR-Cas 3、Cas9、Cas 12 a的两种亚型以及限制性酶HsdRMS和EcoRI。Cas蛋白和限制性酶在整个感染过程中不能接近噬菌体DNA，但是工程化EcoRI在隔室内的重新定位使得能够靶向噬菌体和保护宿主细胞。此外，phi KZ对Cas 13 a-一种靶向RNA的CRISPR-Cas酶敏感-可能是由于噬菌体mRNA定位于细胞质。总的来说，我们建议，巨型假单胞菌逃避广谱的DNA靶向核酸酶通过组装蛋白质屏障周围的基因组。
All viruses require strategies to inhibit or evade the immune pathways of cells that they infect. The viruses that infect bacteria, bacteriophages (phages), must avoid immune pathways that target nucleic acids, such as CRISPR-Cas and restriction-modification systems, to replicate efficiently(1). Here we show that jumbo phage phi KZ segregates its DNA from immunity nucleases of its host, Pseudomonas aeruginosa, by constructing a proteinaceous nucleus-like compartment. phi KZ is resistant to many immunity mechanisms that target DNA in vivo, including two subtypes of CRISPR-Cas3, Cas9, Cas12a and the restriction enzymes HsdRMS and EcoRI. Cas proteins and restriction enzymes are unable to access the phage DNA throughout the infection, but engineering the relocalization of EcoRI inside the compartment enables targeting of the phage and protection of host cells. Moreover, phi KZ is sensitive to Cas13a-a CRISPR-Cas enzyme that targets RNA-probably owing to phage mRNA localizing to the cytoplasm. Collectively, we propose that Pseudomonas jumbo phages evade a broad spectrum of DNA-targeting nucleases through the assembly of a protein barrier around their genome.