RSV glycoprotein and genomic RNA dynamics reveal filament assembly prior to the plasma membrane

• Type of Material: Article
• Authors: Daryll Vanover, Emory University; Daisy V. Smith, Emory University; Emmeline L. Blanchard, Emory University; Eric Alonas, Emory University; Jonathan L. Kirschman, Emory University; Aaron W. Lifland, Georgia Institute of Technology; Chiara Zurla, Emory University; Philip Santangelo, Emory University
• Language: English
• Date: 2017-09-22
• Publisher: Nature Publishing Group: Nature Communications
• Publication Version: Final Published Version
• Copyright Statement: © 2017 The Author(s).
• License: Creative Commons Attribution 4.0 International
• Final Published Version (URL): http://dx.doi.org/10.1038/s41467-017-00732-z
• Title of Journal or Parent Work: Nature Communications
• ISSN: 2041-1723
• Volume: 8
• Issue: 1
• Start Page: 667
• End Page: 667
• Grant/Funding Information: This work was funded by NIH grant R01GM094198 and R01GM114561 (P.J.S.) and by a research partnership between Children’s Healthcare of Atlanta and the Georgia Institute of Technology (A.W.L.).; This material is based upon work supported by the National Science Foundation Graduate Research Fellowship Program under Grant No. DGE-1650044 (E.L.B.).
• Keywords: Virus-host interactions; Viral pathogenesis; Endocytosis; Fluorescence imaging
• Research Categories: Biology, Virology; Biology, Genetics; Engineering, Biomedical

Abstract

The human respiratory syncytial virus G protein plays an important role in the entry and assembly of filamentous virions. Here, we report the use of fluorescently labeled soybean agglutinin to selectively label the respiratory syncytial virus G protein in living cells without disrupting respiratory syncytial virus infectivity or filament formation and allowing for interrogations of respiratory syncytial virus virion assembly. Using this approach, we discovered that plasma membrane-bound respiratory syncytial virus G rapidly recycles from the membrane via clathrin-mediated endocytosis. This event is then followed by the dynamic formation of filamentous and branched respiratory syncytial virus particles, and assembly with genomic ribonucleoproteins and caveolae-associated vesicles prior to re-insertion into the plasma membrane. We demonstrate that these processes are halted by the disruption of microtubules and inhibition of molecular motors. Collectively, our results show that for respiratory syncytial virus assembly, viral filaments are produced and loaded with genomic RNA prior to insertion into the plasma membrane.