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Correspondence: erwright2@wisc.edu or erwrigh@emory.edu; Tel.: +1-608-265-0666; Fax: +1-608-265-4693

Z.K. and E.R.W. conceived of the project and wrote the manuscript.

Z.K., R.S.D., F.L., C.M.H., J.D.S., R.S., M.J. and E.R.W. contributed to cryo-ET imaging and data analysis

Z.K., T.C., F.L., C.C.S., D.R. and C.A.R. contributed to viral growth curve, real time RT-PCR, flow cytometry and infection of polarized NHBE cells, and fusion inhibitor experiment.

Z.K., J.V.T., H.Y. and E.R.W. contributed to the thin section TEM of polarized NHBE cells and imaging.

R.S.P., T.V.H. and M.L.M. contributed the RSV clinical isolate TN strain (A/TN/12/11-19).

Z.K., E.R.W., B.S.G., M.L.M., L.J.A., R.S.P. and T.V.H. contributed to the critical revision of the manuscript.

All authors discussed the results and commented on the manuscript.

We thank Michael Currier, Stacey Human, and Sujin Lee from the Moore Laboratory (Emory University) for the helpful discussion in fusion inhibitor experiments.

We also thank Calvin Cotton (Case Western Reserve University) for providing the NHBE cells.

We would like to thank the Robert P. Apkarian Integrated Electron Microscopy Core of Emory University for microscopy services and support.

Martin L. Moore is an employee of Meissa Vaccines, Inc.

Other authors declare no competing financial interests.

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Research Funding:

This work was supported in part by Emory University, Children’s Healthcare of Atlanta, and the Georgia Research Alliance to E.R.W.; the Center for AIDS Research at Emory University (P30 AI050409); the James B. Pendleton Charitable Trust to E.R.W.; public health service grants R21 AI101775 and R01 GM114561 to E.R.W. from the NIH, and NSF grant 0923395 to E.R.W.

This work was also supported by R01 AI087798 to M.L.M., U19 AI095227 to M.L.M., L.J.A. and R.S.P., R01 AI111820 to R.S.P., United States Department of Veterans Affairs 2I01BX000624 to R.S.P., K24 AI77930 to T.V.H., T32 AI074492 to C.C.S., K12 HD072245 to C.A.R., and F32 GM112517 to J.D.S., R.S. was supported by Emory BCDB Training Grant T32 GM008367-28.

This work was also supported in part by the Emory Children’s Center for Childhood Infections and Vaccines (CCIV).

This work was supported in part by intramural funding from the National Institute of Allergy and Infectious Diseases to B.S.G.

Keywords:

  • Science & Technology
  • Life Sciences & Biomedicine
  • Virology
  • human respiratory syncytial virus (RSV)
  • cryo-electron tomography (cryo-ET)
  • cryo-electron microscopy (cryo-EM)
  • enveloped virus
  • virus assembly
  • viral morphogenesis
  • POLARIZED EPITHELIAL-CELLS
  • NUCLEOPROTEIN-RNA COMPLEX
  • NF-KAPPA-B
  • MATRIX PROTEIN
  • NONSTRUCTURAL PROTEINS
  • FUSION GLYCOPROTEIN
  • M2-1 PROTEIN
  • VACCINE DEVELOPMENT
  • CRYSTAL-STRUCTURE
  • RSV VACCINE

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

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Journal Title:

Viruses

Volume:

Volume 10, Number 8

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Type of Work:

Article | Final Publisher PDF

Abstract:

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.

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© 2018 by the authors.Licensee MDPI, Basel, Switzerland.

This is an Open Access work distributed under the terms of the Creative Commons Attribution 4.0 International License (https://creativecommons.org/licenses/by/4.0/).
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