Department of Biochemistry and Molecular Genetics, University of Alabama at. Birmingham (UAB), 1825 University Blvd, Birmingham, AL, USA.
Department of Chemistry and Chemical Biology, Rutgers, The State University of New Jersey, Piscataway, NJ, USA.
Nat Commun. 2024 Oct 1;15(1):8482. doi: 10.1038/s41467-024-52752-1.
DEV is an obligatory lytic Pseudomonas phage of the N4-like genus, recently reclassified as Schitoviridae. The DEV genome encodes 91 ORFs, including a 3398 amino acid virion-associated RNA polymerase (vRNAP). Here, we describe the complete architecture of DEV, determined using a combination of cryo-electron microscopy localized reconstruction, biochemical methods, and genetic knockouts. We built de novo structures of all capsid factors and tail components involved in host attachment. We demonstrate that DEV long tail fibers are essential for infection of Pseudomonas aeruginosa but dispensable for infecting mutants with a truncated lipopolysaccharide devoid of the O-antigen. We determine that DEV vRNAP is part of a three-gene operon conserved in 191 Schitoviridae genomes. We propose these three proteins are ejected into the host to form a genome ejection motor spanning the cell envelope. We posit that the design principles of the DEV ejection apparatus are conserved in all Schitoviridae.
DEV 是一种必需的裂解假单胞菌噬菌体，属于 N4 样属，最近被重新分类为 Schitoviridae。DEV 基因组编码 91 个 ORF，包括一个 3398 个氨基酸的病毒相关 RNA 聚合酶（vRNAP）。在这里，我们使用冷冻电子显微镜定位重建、生化方法和遗传敲除相结合的方法，描述了 DEV 的完整结构。我们从头构建了所有参与宿主附着的衣壳因子和尾部组件的结构。我们证明 DEV 长尾纤维对于感染铜绿假单胞菌是必不可少的，但对于感染缺乏 O 抗原的截短脂多糖的突变体是可有可无的。我们确定 DEV vRNAP 是在 191 个 Schitoviridae 基因组中保守的三个基因操纵子的一部分。我们提出这三个蛋白被喷射到宿主中，形成跨越细胞膜的基因组喷射马达。我们假设 DEV 喷射装置的设计原则在所有 Schitoviridae 中都是保守的。
