title: Classical swine fever virus Npro interacts with interferon regulatory factor 3 and induces its proteasomal degradation.
abstract: Viruses have evolved a multitude of strategies to subvert the innate immune system by interfering with components of the alpha/beta interferon (IFN-alpha/beta) induction and signaling pathway. It is well established that the pestiviruses prevent IFN-alpha/beta induction in their primary target cells, such as epitheloidal and endothelial cells, macrophages, and conventional dendritic cells, a phenotype mediated by the viral protein N(pro). Central players in the IFN-alpha/beta induction cascade are interferon regulatory factor 3 (IRF3) and IRF7. Recently, it was proposed that classical swine fever virus (CSFV), the porcine pestivirus, induced the loss of IRF3 by inhibiting the transcription of IRF3 mRNA. In the present study, we show that endogenous IRF3 and IRF3 expressed from a cytomegalovirus (CMV) promoter are depleted in the presence of CSFV by means of N(pro), while CSFV does not inhibit CMV promoter-driven protein expression. We also demonstrate that CSFV does not reduce the transcriptional activity of the IRF3 promoter and does not affect the stability of IRF3 mRNA. In fact, CSFV N(pro) induces proteasomal degradation of IRF3, as demonstrated by proteasome inhibition studies. Furthermore, N(pro) coprecipitates with IRF3, suggesting that the proteasomal degradation of IRF3 is induced by a direct or indirect interaction with N(pro). Finally, we show that N(pro) does not downregulate IRF7 expression.
authors.affiliation: Institute of Virology and Immunoprophylaxis (IVI), Sensemattstrasse 293, CH-3147 Mittelhäusern, Switzerland.
authors.forename: Oliver
authors.initials: O
authors.lastname: Bauhofer
authors.forename: Artur
authors.initials: A
authors.lastname: Summerfield
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Sakoda
authors.forename: Jon-Duri
authors.initials: JD
authors.lastname: Tratschin
authors.forename: Martin A
authors.initials: MA
authors.lastname: Hofmann
authors.forename: Nicolas
authors.initials: N
authors.lastname: Ruggli
mesh_terms: D021581:Active Transport, Cell Nucleus; D000818:Animals; D002460:Cell Line; D006692:Classical Swine Fever Virus / Q000235:genetics / Q000378:metabolism*; D003587:Cytomegalovirus / Q000235:genetics; D010450:Endopeptidases / Q000235:genetics / Q000378:metabolism*; D005786:Gene Expression Regulation; D006801:Humans; D050838:Interferon Regulatory Factor-3 / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D010766:Phosphorylation; D011401:Promoter Regions, Genetic / Q000235:genetics; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism*; D061988:Proteasome Inhibitors; D011485:Protein Binding; D012333:RNA, Messenger / Q000235:genetics; D015398:Signal Transduction; D013552:Swine; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D050838:Interferon Regulatory Factor-3; D061988:Proteasome Inhibitors; D012333:RNA, Messenger; D014764:Viral Proteins; D010450:Endopeptidases; C505229:N(pro) protein, swine fever virus; D046988:Proteasome Endopeptidase Complex