The Important Role of Lipid Raft-Mediated Attachment in the Infection of Cultured Cells by Coronavirus Infectious Bronchitis Virus Beaudette Strain

PLoS One. 2017 Jan 12;12(1):e0170123. doi: 10.1371/journal.pone.0170123. eCollection 2017.

Abstract

Lipid raft is an important element for the cellular entry of some viruses, including coronavirus infectious bronchitis virus (IBV). However, the exact role of lipid rafts in the cellular membrane during the entry of IBV into host cells is still unknown. In this study, we biochemically fractionated IBV-infected cells via sucrose density gradient centrifugation after depleting plasma membrane cholesterol with methyl-β-cyclodextrin or Mevastatin. Our results demonstrated that unlike IBV non-structural proteins, IBV structural proteins co-localized with lipid raft marker caveolin-1. Infectivity assay results of Vero cells illustrated that the drug-induced disruption of lipid rafts significantly suppressed IBV infection. Further studies revealed that lipid rafts were not required for IBV genome replication or virion release at later stages. However, the drug-mediated depletion of lipid rafts in Vero cells before IBV attachment significantly reduced the expression of viral structural proteins, suggesting that drug treatment impaired the attachment of IBV to the cell surface. Our results indicated that lipid rafts serve as attachment factors during the early stages of IBV infection, especially during the attachment stage.

MeSH terms

  • A549 Cells
  • Animals
  • Caveolin 1 / metabolism
  • Cell Membrane / drug effects
  • Cell Membrane / metabolism
  • Chlorocebus aethiops
  • Cholesterol / metabolism
  • Endocytosis
  • Humans
  • Infectious bronchitis virus / genetics
  • Infectious bronchitis virus / physiology*
  • Membrane Microdomains / drug effects
  • Membrane Microdomains / metabolism*
  • Membrane Microdomains / virology
  • Microscopy, Fluorescence
  • RNA, Viral / analysis
  • Real-Time Polymerase Chain Reaction
  • Vero Cells
  • Viral Envelope Proteins / metabolism
  • Virus Replication
  • beta-Cyclodextrins / pharmacology

Substances

  • Caveolin 1
  • RNA, Viral
  • Viral Envelope Proteins
  • beta-Cyclodextrins
  • methyl-beta-cyclodextrin
  • Cholesterol

Grants and funding

This research was supported by grants from International Science & Technology Cooperation Program of China (2014DFA31890), the National Natural Science Foundation of China (31672592, 31602038, 31602083), Elite Youth Program of Chinese Academy of Agricultural Sciences, China, and the National Science and Technology Support Program (2013BAD12B00).