Clathrin- and serine proteases-dependent uptake of porcine epidemic diarrhea virus into Vero cells

Virus Res. 2014 Oct 13;191:21-9. doi: 10.1016/j.virusres.2014.07.022. Epub 2014 Jul 31.

Abstract

Porcine epidemic diarrhea virus (PEDV), a member of the genus Alphacoronavirus, is a causative agent of porcine enteric disease characterized by acute watery diarrhea and dehydration in sucking piglet. Similar to other coronaviruses, PEDV spike protein mediates its cell entry by binding to cellular receptors and inducing membrane fusion between viral envelopes and cellular membranes. However, the entry mechanism of PEDV is not studied. Here, we determined the entry mechanism of PEDV into Vero cells. Our data confirmed that PEDV entry followed clathrin-mediated endocytosis independence of caveolae-coated pit assembly. The internalized PEDV was co-localized with the clathrin-mediated endocytic marker, but not with the caveolae-mediated endocytic marker. In addition, cells treated with lysosomotropic agents and serine protease inhibitors were resistant to PEDV. Our data revealed that PEDV entry followed clathrin-mediated endocytosis and was dependent on a low pH and serine proteolysis for successful entry into cells.

Keywords: Clathrin; Coronavirus; Entry; Low pH; Porcine epidemic diarrhea; Serine protease.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Chlorocebus aethiops
  • Clathrin / metabolism*
  • Coronavirus Infections / enzymology
  • Coronavirus Infections / metabolism
  • Coronavirus Infections / veterinary*
  • Coronavirus Infections / virology
  • Endocytosis
  • Porcine epidemic diarrhea virus / physiology*
  • Serine Endopeptidases / metabolism*
  • Swine
  • Swine Diseases / enzymology*
  • Swine Diseases / metabolism
  • Swine Diseases / virology
  • Vero Cells
  • Virus Internalization*

Substances

  • Clathrin
  • Serine Endopeptidases