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

• Published in: Viruses Vol. 10; no. 8; p. 446
• Main Authors: Ke, Zunlong, Dillard, Rebecca S, Chirkova, Tatiana, Leon, Fredrick, Stobart, Christopher C, Hampton, Cheri M, Strauss, Joshua D, Rajan, Devi, Rostad, Christina A, Taylor, Jeannette V, Yi, Hong, Shah, Raven, Jin, Mengtian, Hartert, Tina V, Peebles, Jr, R Stokes, Graham, Barney S, Moore, Martin L, Anderson, Larry J, Wright, Elizabeth R
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 20.08.2018; MDPI
• Subjects: A549 Cells; Animals; Bronchi - virology; Cell Line; Cell lines; Cercopithecus aethiops; Children; cryo-electron microscopy (cryo-EM); cryo-electron tomography (cryo-ET); Cryoelectron Microscopy - methods; Electron Microscope Tomography - methods; enveloped virus; Epithelial Cells - ultrastructure; Epithelial Cells - virology; Geriatrics; Glycoproteins; HeLa Cells; human respiratory syncytial virus (RSV); Humans; Infectious diseases; Laboratories; Measles; Microscopy; Microtomy; Morphogenesis; Morphology; Mutation; Pneumoviridae; Proteins; Respiratory syncytial virus; Respiratory Syncytial Virus, Human - physiology; Respiratory Syncytial Virus, Human - ultrastructure; Respiratory tract diseases; RNA polymerase; RNA viruses; Studies; Tomography; Transmission electron microscopy; Vaccines; Vero Cells; viral morphogenesis; Virion - physiology; Virion - ultrastructure; virus assembly; Virus Assembly - physiology; virus assembly; human respiratory syncytial virus (RSV); cryo-electron tomography (cryo-ET); cryo-electron microscopy (cryo-EM); enveloped virus; viral morphogenesis
• ISSN: 1999-4915; 1999-4915
• DOI: 10.3390/v10080446

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 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.