Impairing the function of MLCK, myosin Va or myosin Vb disrupts Rhinovirus B14 replication

Sci Rep. 2017 Dec 7;7(1):17153. doi: 10.1038/s41598-017-17501-z.

Abstract

Together, the three human rhinovirus (RV) species are the most frequent cause of the common cold. Because of their high similarity with other viral species of the genus Enterovirus, within the large family Picornaviridae, studies on RV infectious activities often offer a less pathogenic model for more aggressive enteroviruses, e.g. poliovirus or EV71. Picornaviruses enter via receptor mediated endocytosis and replicate in the cytosol. Most of them depend on functional F-actin, Rab proteins, and probably motor proteins. To assess the latter, we evaluated the role of myosin light chain kinase (MLCK) and two myosin V isoforms (Va and Vb) in RV-B14 infection. We report that ML-9, a very specific MLCK inhibitor, dramatically reduced RV-B14 entry. We also demonstrate that RV-B14 infection in cells expressing dominant-negative forms of myosin Va and Vb was impaired after virus entry. Using immunofluorescent localization and immunoprecipitation, we show that myosin Va co-localized with RV-B14 exclusively after viral entry (15 min post infection) and that myosin Vb was present in the clusters of newly synthesized RNA in infected cells. These clusters, observed at 180 min post infection, are reminiscent of replication sites. Taken together, these results identify myosin light chain kinase, myosin Va and myosin Vb as new players in RV-B14 infection that participate directly or indirectly in different stages of the viral cycle.

MeSH terms

  • Actin Cytoskeleton / metabolism
  • Enterovirus Infections / metabolism
  • Enterovirus Infections / prevention & control*
  • Enterovirus Infections / virology
  • HeLa Cells
  • Humans
  • Methylmalonic Acid / analogs & derivatives*
  • Methylmalonic Acid / pharmacology
  • Myosin Heavy Chains / antagonists & inhibitors*
  • Myosin Type V / antagonists & inhibitors*
  • Myosin-Light-Chain Kinase / antagonists & inhibitors*
  • Phosphorylation
  • Rhinovirus / drug effects
  • Rhinovirus / physiology
  • Virus Internalization / drug effects*
  • Virus Replication / drug effects*

Substances

  • MYO5B protein, human
  • butyl-2-methyl-malonic acid
  • MYO5A protein, human
  • Methylmalonic Acid
  • Myosin-Light-Chain Kinase
  • Myosin Type V
  • Myosin Heavy Chains