The Morphology and Assembly of Respiratory Syncytial Virus Revealed by Cryo-Electron Tomography

Viruses

TinaV. Hartert¹  R.Stokes Peebles¹  Tatiana Chirkova²  Zunlong Ke²  ChristinaA. Rostad²  RebeccaS. Dillard²  ChristopherC. Stobart²  CheriM. Hampton²  Fredrick Leon²  Raven Shah²  Mengtian Jin²  LarryJ. Anderson²  ElizabethR. Wright²  MartinL. Moore²  Devi Rajan²  JoshuaD. Strauss²  Hong Yi³  JeannetteV. Taylor³  BarneyS. Graham⁴ 

[1] Division of Allergy, Pulmonary, and Critical Care Medicine, Vanderbilt University School of Medicine, Nashville, TN 37232, USA;Division of Pediatric Infectious Diseases, Emory University School of Medicine, Children’s Healthcare of Atlanta, Atlanta, GA 30322, USA;Robert P. Apkarian Integrated Electron Microscopy Core, Emory University, Atlanta, GA 30322, USA;Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892, USA;

关键词: human respiratory syncytial virus (RSV);    cryo-electron tomography (cryo-ET);    cryo-electron microscopy (cryo-EM);    enveloped virus;    virus assembly;    viral morphogenesis;   

DOI  :  10.3390/v10080446

来源: DOAJ

【 摘 要 】

Human respiratory syncytial virus (RSV) is the leading cause of lower respiratory tract disease in young children. With repeat infections throughout life, it can also cause substantial disease in the elderly and in adults with compromised cardiac, pulmonary and immune systems. RSV is a pleomorphic enveloped RNA virus in the Pneumoviridae family. Recently, the three-dimensional (3D) structure of purified RSV particles has been elucidated, revealing three distinct morphological categories: spherical, asymmetric, and filamentous. However, the native 3D structure of RSV particles associated with or released from infected cells has yet to be investigated. In this study, we have established an optimized system for studying RSV structure by imaging RSV-infected cells on transmission electron microscopy (TEM) grids by cryo-electron tomography (cryo-ET). Our results demonstrate that RSV is filamentous across several virus strains and cell lines by cryo-ET, cryo-immuno EM, and thin section TEM techniques. The viral filament length varies from 0.5 to 12 μm and the average filament diameter is approximately 130 nm. Taking advantage of the whole cell tomography technique, we have resolved various stages of RSV assembly. Collectively, our results can facilitate the understanding of viral morphogenesis in RSV and other pleomorphic enveloped viruses.

【 授权许可】

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