Coronavirus S protein-induced fusion is blocked prior to hemifusion by Abl kinase inhibitors

J Gen Virol. 2018 May;99(5):619-630. doi: 10.1099/jgv.0.001047. Epub 2018 Mar 20.

Abstract

Enveloped viruses gain entry into host cells by fusing with cellular membranes, a step that is required for virus replication. Coronaviruses, including the severe acute respiratory syndrome coronavirus (SARS-CoV), Middle East respiratory syndrome coronavirus (MERS-CoV) and infectious bronchitis virus (IBV), fuse at the plasma membrane or use receptor-mediated endocytosis and fuse with endosomes, depending on the cell or tissue type. The virus spike (S) protein mediates fusion with the host cell membrane. We have shown previously that an Abelson (Abl) kinase inhibitor, imatinib, significantly reduces SARS-CoV and MERS-CoV viral titres and prevents endosomal entry by HIV SARS S and MERS S pseudotyped virions. SARS-CoV and MERS-CoV are classified as BSL-3 viruses, which makes experimentation into the cellular mechanisms involved in infection more challenging. Here, we use IBV, a BSL-2 virus, as a model for studying the role of Abl kinase activity during coronavirus infection. We found that imatinib and two specific Abl kinase inhibitors, GNF2 and GNF5, reduce IBV titres by blocking the first round of virus infection. Additionally, all three drugs prevented IBV S-induced syncytia formation prior to the hemifusion step. Our results indicate that membrane fusion (both virus-cell and cell-cell) is blocked in the presence of Abl kinase inhibitors. Studying the effects of Abl kinase inhibitors on IBV will be useful in identifying the host cell pathways required for coronavirus infection. This will provide an insight into possible therapeutic targets to treat infections by current as well as newly emerging coronaviruses.

Keywords: Abl kinase; Abl1; Abl2; GNF2; GNF5; IBV; MERS-CoV; SARS-CoV; cell-cell fusion; coronavirus; imatinib; virus-cell fusion.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Antiviral Agents / pharmacology
  • Benzamides / pharmacology
  • Cell Membrane
  • Chlorocebus aethiops
  • Endosomes / virology*
  • Imatinib Mesylate / pharmacology
  • Infectious bronchitis virus / genetics*
  • Infectious bronchitis virus / metabolism
  • Protein Kinase Inhibitors / pharmacology*
  • Proto-Oncogene Proteins c-abl / antagonists & inhibitors*
  • Pyrimidines / pharmacology
  • Spike Glycoprotein, Coronavirus / genetics
  • Spike Glycoprotein, Coronavirus / metabolism*
  • Vero Cells
  • Virus Internalization*
  • Virus Replication

Substances

  • Antiviral Agents
  • Benzamides
  • GNF-2 compound
  • N-(2-hydroxyethyl)-3-(6-((4-(trifluoromethoxy)phenyl)amino)-4-pyrimidinyl)benzamide
  • Protein Kinase Inhibitors
  • Pyrimidines
  • Spike Glycoprotein, Coronavirus
  • Imatinib Mesylate
  • Proto-Oncogene Proteins c-abl