In the present study, we investigated the molecular mechanisms by which NS4B targets RIG-I–induced and STING-mediated IFN-β production signaling. IFN-β promoter reporter assay showed that IFN-β promoter activation induced by RIG-I or Cardif was significantly suppressed by both NS4B and NS3/4A, whereas STING-induced IFN-β activation was suppressed by NS4B but not by NS3/4A, suggesting that NS4B had a distinct point of interaction. Immunostaining showed that STING colocalized with NS4B in the endoplasmic reticulum.

Immunoprecipitation and bimolecular fluorescence complementation (BiFC) assays demonstrated that NS4B specifically bound STING. Intriguingly, NS4B expression selleck blocked the protein interaction between STING and Cardif, which is required for robust IFN-β activation. NS4B truncation assays showed that its N terminus, containing the STING homology domain, was necessary for the suppression of IFN-β promoter activation. NS4B suppressed residual IFN-β activation by an NS3/4A-cleaved Cardif (Cardif1-508), suggesting that NS3/4A and NS4B may cooperate in the blockade of IFN-β production. Conclusion:

NS4B suppresses RIG-I–mediated IFN-β production signaling through a direct protein interaction with STING. Disruption of that interaction may restore cellular antiviral responses and may constitute a novel therapeutic strategy for the eradication of HCV. (HEPATOLOGY 2013) Type I interferon (IFN) plays a central role in eliminating hepatitis C virus (HCV) both under physiological conditions and when used as a therapeutic intervention.1-3 In experimental acute-resolving HCV infection in chimpanzees, MCE numerous Small molecule library cell line IFN-related genes are expressed during clinical course of infection.4 Viruses are recognized by cellular innate

immune receptors, such as toll-like receptors, and a family of RIG-I–like receptors, such as retinoic-acid-inducible gene I (RIG-I) and melanoma-differentiation-associated gene 5 (MDA-5); host antiviral responses are then activated, resulting in the production of cytokines such as type I and type III IFNs.5 RIG-I is activated through recognition of short double-strand RNA (dsRNA) or triphosphate at the 5′ end of dsRNA as pathogen-associated molecular patterns,6, 7 forming a homo-oligomer that binds with the caspase recruitment domain (CARD) of Cardif (also known as MAVS, VISA, or IPS-1).8-11 Cardif subsequently recruits TANK binding kinase 1 (TBK1) and IκB kinase ϵ (IKKϵ) kinases, which catalyze phosphorylation and activation of IFN regulatory factor-3 (IRF-3).12 Activation of TBK1 and IKKϵ results in the phosphorylation of IRF-3 or IRF-7, translocation to the nucleus, and induction of IFN-β mRNA transcription. Several HCV proteins can block host cellular antiviral responses. HCV core protein blocks IFN signaling by interacting with signal transducer and activator of transcription protein-1 (STAT1).