Alice-Roza Eruera, James Hodgkinson-Bean, Georgia L Rutter, Francesca R Hills, Rosheny Kumaran, Alexander J M Crowe, Nickhil Jadav, Fangfang Chang, Klemens McJarrow-Keller, Fátima Jorge, Jaekyung Hyun, Hyejin Kim, Bumhan Ryu, Mihnea Bostina
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The core consists of tetrameric and octameric ejection proteins which form a ∼1.5-MDa ejectosome that must transition through the ∼30 Å aperture created by the short tail nozzle assembly that acts as the conduit for the passage of DNA during infection. The ejectosome forms several grooves into which coils of genomic DNA are fit before the DNA sharply turns and goes down the tunnel and into the portal. In addition, we reconstructed the icosahedral capsid and hybrid tail apparatus to resolutions between 3.04 and 3.23 Å, and note an uncommon fold adopted by the dimerized decoration proteins which further emphasize the structural diversity of podophages. These reconstructions have allowed the generation of a complete atomic model of the ΦM1, uncovering two distinct decoration proteins and highlighting the exquisite structural diversity of tailed bacteriophages.</p>","PeriodicalId":74468,"journal":{"name":"PNAS nexus","volume":null,"pages":null},"PeriodicalIF":2.2000,"publicationDate":"2024-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11440229/pdf/","citationCount":"0","resultStr":"{\"title\":\"Ejectosome of <i>Pectobacterium</i> bacteriophage ΦM1.\",\"authors\":\"Alice-Roza Eruera, James Hodgkinson-Bean, Georgia L Rutter, Francesca R Hills, Rosheny Kumaran, Alexander J M Crowe, Nickhil Jadav, Fangfang Chang, Klemens McJarrow-Keller, Fátima Jorge, Jaekyung Hyun, Hyejin Kim, Bumhan Ryu, Mihnea Bostina\",\"doi\":\"10.1093/pnasnexus/pgae416\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Podophages that infect gram-negative bacteria, such as <i>Pectobacterium</i> pathogen ΦM1, encode tail assemblies too short to extend across the complex gram-negative cell wall. 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In addition, we reconstructed the icosahedral capsid and hybrid tail apparatus to resolutions between 3.04 and 3.23 Å, and note an uncommon fold adopted by the dimerized decoration proteins which further emphasize the structural diversity of podophages. 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引用次数: 0
摘要
感染革兰氏阴性细菌(如果胶杆菌病原体ΦM1)的荚膜病毒编码的尾部组件太短,无法穿过复杂的革兰氏阴性细胞壁。为了克服这一问题,荚膜病毒编码了一种大型蛋白质复合物(弹射体),将其包装在病毒外壳内,并在感染过程中相应地弹射出来,形成一个横跨周质空间的瞬时通道。在这里,我们通过单粒子冷冻电镜(cryo-EM)以 3.32 Å 的分辨率描述了噬菌体 ΦM1 的喷射体。核心由四聚体和八聚体喷射蛋白组成,它们形成一个 1.5 兆焦耳的喷射体,喷射体必须穿过由短尾喷嘴组件形成的 30 Å 孔径,短尾喷嘴组件在感染过程中充当 DNA 的通道。喷射体形成几个凹槽,将基因组 DNA 线圈装入其中,然后 DNA 急转弯,沿着隧道进入入口。此外,我们还以 3.04 至 3.23 Å 的分辨率重建了二十面体荚膜和混合尾部装置,并注意到二聚化装饰蛋白采用了一种不常见的折叠方式,这进一步强调了荚膜病毒结构的多样性。通过这些重建,生成了一个完整的ΦM1原子模型,发现了两种不同的装饰蛋白,突出了噬菌体尾部结构的精致多样性。
Podophages that infect gram-negative bacteria, such as Pectobacterium pathogen ΦM1, encode tail assemblies too short to extend across the complex gram-negative cell wall. To overcome this, podophages encode a large protein complex (ejectosome) packaged inside the viral capsid and correspondingly ejected during infection to form a transient channel that spans the periplasmic space. Here, we describe the ejectosome of bacteriophage ΦM1 to a resolution of 3.32 Å by single-particle cryo-electron microscopy (cryo-EM). The core consists of tetrameric and octameric ejection proteins which form a ∼1.5-MDa ejectosome that must transition through the ∼30 Å aperture created by the short tail nozzle assembly that acts as the conduit for the passage of DNA during infection. The ejectosome forms several grooves into which coils of genomic DNA are fit before the DNA sharply turns and goes down the tunnel and into the portal. In addition, we reconstructed the icosahedral capsid and hybrid tail apparatus to resolutions between 3.04 and 3.23 Å, and note an uncommon fold adopted by the dimerized decoration proteins which further emphasize the structural diversity of podophages. These reconstructions have allowed the generation of a complete atomic model of the ΦM1, uncovering two distinct decoration proteins and highlighting the exquisite structural diversity of tailed bacteriophages.