title: Response of Three Different Viruses to Interferon Priming and Dithiothreitol Treatment of Avian Cells.
abstract: We have previously shown that the replication of avian reovirus (ARV) in chicken cells is much more resistant to interferon (IFN) than the replication of vesicular stomatitis virus (VSV) or vaccinia virus (VV). In this study, we have investigated the role that the double-stranded RNA (dsRNA)-activated protein kinase (PKR) plays in the sensitivity of these three viruses toward the antiviral action of chicken interferon. Our data suggest that while interferon priming of avian cells blocks vaccinia virus replication by promoting PKR activation, the replication of vesicular stomatitis virus appears to be blocked at a pretranslational step. Our data further suggest that the replication of avian reovirus in chicken cells is quite resistant to interferon priming because this virus uses strategies to downregulate PKR activation and also because translation of avian reovirus mRNAs is more resistant to phosphorylation of the alpha subunit of initiation factor eIF2 than translation of their cellular counterparts. Our results further reveal that the avian reovirus protein sigmaA is able to prevent PKR activation and that this function is dependent on its double-stranded RNA-binding activity. Finally, this study demonstrates that vaccinia virus and avian reovirus, but not vesicular stomatitis virus, express/induce factors that counteract the ability of dithiothreitol to promote eIF2 phosphorylation. Our data demonstrate that each of the three different viruses used in this study elicits distinct responses to interferon and to dithiothreitol-induced eIF2 phosphorylation when infecting avian cells.
  
  
  
  Type I interferons constitute the first barrier of defense against viral infections, and one of the best characterized antiviral strategies is mediated by the double-stranded RNA-activated protein kinase R (PKR). The results of this study revealed that IFN priming of avian cells has little effect on avian reovirus (ARV) replication but drastically diminishes the replication of vaccinia virus (VV) and vesicular stomatitis virus (VSV) by PKR-dependent and -independent mechanisms, respectively. Our data also demonstrate that the dsRNA-binding ability of ARV protein sigmaA plays a key role in the resistance of ARV toward IFN by preventing PKR activation. Our findings will contribute to improve the current understanding of the interaction of viruses with the host's innate immune system. Finally, it would be of interest to uncover the mechanisms that allow avian reovirus transcripts to be efficiently translated under conditions (moderate eIF2 phosphorylation) that block the synthesis of cellular proteins.
authors.affiliation: Centro Singular de Investigación en Química Biolóxica e Materiais Moleculares and Departamento de Bioquímica e Bioloxía Molecular, Universidade de Santiago de Compostela, Santiago de Compostela, Spain.
authors.forename: Irene
authors.initials: I
authors.lastname: Lostalé-Seijo
authors.affiliation: Centro Singular de Investigación en Química Biolóxica e Materiais Moleculares and Departamento de Bioquímica e Bioloxía Molecular, Universidade de Santiago de Compostela, Santiago de Compostela, Spain.
authors.forename: José
authors.initials: J
authors.lastname: Martínez-Costas
authors.affiliation: Centro Singular de Investigación en Química Biolóxica e Materiais Moleculares and Departamento de Bioquímica e Bioloxía Molecular, Universidade de Santiago de Compostela, Santiago de Compostela, Spain franciscojavier.benavente@usc.es.
authors.forename: Javier
authors.initials: J
authors.lastname: Benavente
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000378:metabolism*; D002460:Cell Line; D002645:Chickens; D004229:Dithiothreitol / Q000378:metabolism; D015852:Eukaryotic Initiation Factor-2 / Q000378:metabolism; D054884:Host-Pathogen Interactions; D007372:Interferons / Q000378:metabolism*; D030101:Orthoreovirus, Avian / Q000502:physiology*; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D014616:Vaccinia virus / Q000502:physiology*; D018116:Vesiculovirus / Q000502:physiology*; D014779:Virus Replication / Q000187:drug effects*; D019892:eIF-2 Kinase / Q000378:metabolism*
chemical_list: D000998:Antiviral Agents; D015852:Eukaryotic Initiation Factor-2; D007372:Interferons; D019892:eIF-2 Kinase; D004229:Dithiothreitol