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A bacteriophage nucleus-like compartment shields DNA from CRISPR nucleases

机译：噬菌体核样隔室可屏蔽CRISPR核酸酶的DNA

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摘要

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 FKZ segregates its DNA from immunity nucleases of its host, Pseudomonas aeruginosa, by constructing a proteinaceous nucleus-like compartment. FKZ 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.
机译：所有病毒都需要采取策略来抑制或逃避其感染细胞的免疫途径。感染细菌，噬菌体（噬菌体）的病毒必须避免针对核酸的免疫途径，例如CRISPR-Cas和限制性修饰系统才能有效复制（1）。在这里我们显示，巨噬细胞FKZ通过构建蛋白核样区室，将其DNA与宿主铜绿假单胞菌的免疫核酸酶分离。 FKZ对体内靶向DNA的许多免疫机制具有抵抗力，包括CRISPR-Cas3的两种亚型，Cas9，Cas12a和限制性酶HsdRMS和EcoRI。 Cas蛋白和限制性内切酶在整个感染过程中均无法访问噬菌体DNA，但通过对EcoRI在隔室内进行重新定位，可以靶向噬菌体并保护宿主细胞。此外，Phi KZ对Cas13a-一种CRISPR-Cas酶敏感，该酶靶向RNA，很可能是由于噬菌体mRNA定位于细胞质。总体而言，我们建议假单胞菌巨型噬菌体通过在其基因组周围组装蛋白质屏障来逃避广谱的靶向DNA的核酸酶。

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  • 来源
    《Nature》 |2020年第7789期|244-248|共5页
  • 作者

    Mendoza Senen D.; Nieweglowska Eliza S.; Govindarajan Sutharsan; Leon Lina M.; Berry Joel D.; Tiwari Anika; Chaikeeratisak Vorrapon; Pogliano Joe; Agard David A.; Bondy-Denomy Joseph;

  • 作者单位

    Univ Calif San Francisco Dept Microbiol & Immunol San Francisco CA 94143 USA;

    Univ Calif San Francisco Dept Biochem & Biophys Howard Hughes Med Inst San Francisco CA 94143 USA;

    Univ Calif San Francisco Dept Microbiol & Immunol San Francisco CA 94143 USA|SRM Univ AP Dept Biol Amaravati India;

    Univ Calif San Diego Div Biol Sci La Jolla CA 92093 USA|Chulalongkorn Univ Fac Sci Dept Biochem Bangkok Thailand;

    Univ Calif San Diego Div Biol Sci La Jolla CA 92093 USA;

    Univ Calif San Francisco Dept Biochem & Biophys Howard Hughes Med Inst San Francisco CA 94143 USA|Univ Calif San Francisco Quantitat Biosci Inst San Francisco CA 94143 USA;

    Univ Calif San Francisco Dept Microbiol & Immunol San Francisco CA 94143 USA|Univ Calif San Francisco Quantitat Biosci Inst San Francisco CA 94143 USA;

  • 收录信息 美国《科学引文索引》(SCI);美国《工程索引》(EI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
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