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

Nat Commun. 2017 Sep 22;8(1):667. doi: 10.1038/s41467-017-00732-z.

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.Assembly of filamentous RSV particles is incompletely understood due to a lack of techniques suitable for live-cell imaging. Here Vanover et al. use labeled soybean agglutinin to selectively label RSV G protein and show how filamentous RSV assembly, initiated in the cytoplasm, uses G protein recycled from the plasma membrane.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Caveolins / metabolism
  • Cell Membrane / metabolism*
  • Chlorocebus aethiops
  • Clathrin / metabolism
  • Endocytosis / physiology
  • Humans
  • Microtubules / metabolism
  • Molecular Imaging / methods
  • Plant Lectins / chemistry
  • Plant Lectins / metabolism
  • RNA, Viral / metabolism*
  • Respiratory Syncytial Virus, Human / genetics*
  • Respiratory Syncytial Virus, Human / pathogenicity
  • Respiratory Syncytial Virus, Human / physiology
  • Ribonucleoproteins / metabolism
  • Soybean Proteins / chemistry
  • Soybean Proteins / metabolism
  • Vero Cells
  • Viral Fusion Proteins / metabolism*
  • Virus Replication

Substances

  • Caveolins
  • Clathrin
  • G glycoprotein, Respiratory syncytial virus
  • Plant Lectins
  • RNA, Viral
  • Ribonucleoproteins
  • Soybean Proteins
  • Viral Fusion Proteins
  • soybean lectin