title: Hepatitis C virus NS2 protease inhibits host cell antiviral response by inhibiting IKKε and TBK1 functions.
abstract: Hepatitis C virus (HCV) encodes for several proteins that can interfere with host cell signaling and antiviral response. Previously, serine protease NS3/4A was shown to block host cell interferon (IFN) production by proteolytic cleavage of MAVS and TRIF, the adaptor molecules of the RIG-I and TLR3 signaling pathways, respectively. This study shows that another HCV protease, NS2 can interfere efficiently with cytokine gene expression. NS2 and its proteolytically inactive mutant forms were able to inhibit type I and type III IFN, CCL5 and CXCL10 gene promoters activated by Sendai virus infection. However, the CXCL8 gene promoter was not inhibited by NS2. In addition, constitutively active RIG-I (ΔRIG-I), MAVS, TRIF, IKKε, and TBK1-induced activation of IFN-β promoter was inhibited by NS2. Cotransfection experiments with IKKε or TBK1 together with interferon regulatory factor 3 (IRF3) and HCV expression constructs revealed that NS2 in a dose-dependent manner inhibited IKKε and especially TBK1-induced IRF3 phosphorylation. GST pull-down experiments with GST-NS2 and in vitro-translated and cell-expressed IKKε and TBK1 demonstrated direct physical interactions of the kinases with NS2. Further evidence that the IKKε/TBK1 kinase complex is the target for NS2 was obtained from the observation that the constitutively active form of IRF3 (IRF3-5D) activated readily IFN-β promoter in the presence of NS2. The present study identified HCV NS2 as a potent interferon antagonist, and describes an explanation of how NS2 downregulates the major signaling pathways involved in the development of host innate antiviral responses.
authors.affiliation: Virology Unit, Department of Infectious Disease Surveillance and Control, National Institute for Health and Welfare (THL), Helsinki, Finland. pasi.kaukinen@helsinki.fi
authors.forename: Pasi
authors.initials: P
authors.lastname: Kaukinen
authors.forename: Maarit
authors.initials: M
authors.lastname: Sillanpää
authors.forename: Laura
authors.initials: L
authors.lastname: Nousiainen
authors.forename: Krister
authors.initials: K
authors.lastname: Melén
authors.forename: Ilkka
authors.initials: I
authors.lastname: Julkunen
mesh_terms: D002498:Centrifugation; D016207:Cytokines / Q000037:antagonists & inhibitors; D016174:Hepacivirus / Q000276:immunology / Q000472:pathogenicity*; D054884:Host-Pathogen Interactions; D006801:Humans; D051550:I-kappa B Kinase / Q000037:antagonists & inhibitors* / Q000276:immunology; D011485:Protein Binding; D025941:Protein Interaction Mapping; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors* / Q000276:immunology; D017361:Viral Nonstructural Proteins / Q000276:immunology / Q000378:metabolism*
chemical_list: D016207:Cytokines; C097010:NS2 protein, Hepatitis C virus; D017361:Viral Nonstructural Proteins; D017346:Protein Serine-Threonine Kinases; C403191:TBK1 protein, human; D051550:I-kappa B Kinase