title: Orf virus inhibits interferon stimulated gene expression and modulates the JAK/STAT signalling pathway.
abstract: Interferons (IFNs) play a critical role as a first line of defence against viral infection. Activation of the Janus kinase/signal transducer and activation of transcription (JAK/STAT) pathway by IFNs leads to the production of IFN stimulated genes (ISGs) that block viral replication. The Parapoxvirus, Orf virus (ORFV) induces acute pustular skin lesions of sheep and goats and is transmissible to man. The virus replicates in keratinocytes that are the immune sentinels of skin. We investigated whether or not ORFV could block the expression of ISGs. The human gene GBP1 is stimulated exclusively by type II IFN while MxA is stimulated exclusively in response to type I IFNs. We found that GBP1 and MxA were strongly inhibited in ORFV infected HeLa cells stimulated with IFN-γ or IFN-α respectively. Furthermore we showed that ORFV inhibition of ISG expression was not affected by cells pretreated with adenosine N1-oxide (ANO), a molecule that inhibits poxvirus mRNA translation. This suggested that new viral gene synthesis was not required and that a virion structural protein was involved. We next investigated whether ORFV infection affected STAT1 phosphorylation in IFN-γ or IFN-α treated HeLa cells. We found that ORFV reduced the levels of phosphorylated STAT1 in a dose-dependent manner and was specific for Tyr701 but not Ser727. Treatment of cells with sodium vanadate suggested that a tyrosine phosphatase was responsible for dephosphorylating STAT1-p. ORFV encodes a factor, ORFV057, with homology to the vaccinia virus structural protein VH1 that impairs the JAK/STAT pathway by dephosphorylating STAT1. Our findings show that ORFV has the capability to block ISG expression and modulate the JAK/STAT signalling pathway.
authors.affiliation: Virus Research Unit, Department of Microbiology and Immunology, University of Otago, New Zealand. Electronic address: ryanpaulharvey@gmail.com.
authors.forename: Ryan
authors.initials: R
authors.lastname: Harvey
authors.affiliation: Virus Research Unit, Department of Microbiology and Immunology, University of Otago, New Zealand. Electronic address: catherine.mccaughan@otago.ac.nz.
authors.forename: Catherine
authors.initials: C
authors.lastname: McCaughan
authors.affiliation: Virus Research Unit, Department of Microbiology and Immunology, University of Otago, New Zealand. Electronic address: lyn.wise@otago.ac.nz.
authors.forename: Lyn M
authors.initials: LM
authors.lastname: Wise
authors.affiliation: Virus Research Unit, Department of Microbiology and Immunology, University of Otago, New Zealand. Electronic address: andy.mercer@otago.ac.nz.
authors.forename: Andrew A
authors.initials: AA
authors.lastname: Mercer
authors.affiliation: Virus Research Unit, Department of Microbiology and Immunology, University of Otago, New Zealand. Electronic address: stephen.fleming@otago.ac.nz.
authors.forename: Stephen B
authors.initials: SB
authors.lastname: Fleming
mesh_terms: D002460:Cell Line; D019204:GTP-Binding Proteins / Q000378:metabolism; D006367:HeLa Cells; D054884:Host-Pathogen Interactions; D006801:Humans; D007372:Interferons / Q000378:metabolism*; D053612:Janus Kinases / Q000235:genetics / Q000378:metabolism*; D064547:Myxovirus Resistance Proteins / Q000378:metabolism; D009923:Orf virus / Q000235:genetics / Q000378:metabolism; D010766:Phosphorylation; D011213:Poxviridae Infections / Q000235:genetics* / Q000378:metabolism; D050794:STAT1 Transcription Factor / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: C490287:GBP1 protein, human; C579071:MX1 protein, human; D064547:Myxovirus Resistance Proteins; D050794:STAT1 Transcription Factor; C494079:STAT1 protein, human; D014764:Viral Proteins; D007372:Interferons; D053612:Janus Kinases; D019204:GTP-Binding Proteins
keywords: Immuno-modulation; Interferon; JAK/STAT signalling pathway; Orf virus; Parapoxvirus