Molecular Cloning, Characterization and Expression Analysis of Woodchuck Retinoic Acid-Inducible Gen

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Cytosolic retinoic acid-inducible gene I(RIG-I) is an important innate immune RNA sensor and can induce antiviral cytokines, e.g., interferon-β(IFN-β). Innate immune response to hepatitis B virus(HBV) plays a pivotal role in viral clearance and persistence. However, knowledge of the role that RIG-I plays in HBV infection is limited. The woodchuck is a valuable model for studying HBV infection. To characterize the molecular basis of woodchuck RIG-I(w RIG-I), we analyzed the complete coding sequences(CDSs) of w RIG-I, containing 2778 base pairs that encode 925 amino acids. The deduced w RIG-I protein was 106.847 k D with a theoretical isoelectric point(p I) of 6.07, and contained three important functional structures [caspase activation and recruitment domains(CARDs), DEx D/H-box helicases, and a repressor domain(RD)]. In woodchuck fibroblastoma cell line(WH12/6), w RIG-I-targeted small interfering RNA(si RNA) down-regulated RIG-I and its downstrean effector–IFN-β transcripts under RIG-I’ ligand, 5’-ppp double stranded RNA(ds RNA) stimulation. We also measured m RNA levels of w RIG-I in different tissues from healthy woodchucks and in the livers from woodchuck hepatitis virus(WHV)-infected woodchucks. The basal expression levels of w RIG-I were abundant in the kidney and liver. Importantly, w RIG-I was significantly up-regulated in acutely infected woodchuck livers, suggesting that RIG-I might be involved in WHV infection. These results may characterize RIG-I in the woodchuck model, providing a strong basis for further study on RIG-I-mediated innate immunity in HBV infection. Innate immune response to hepatitis B virus (HBV) plays an pivotal role (RIG-I) is an important innate immune RNA sensor and can induce antiviral cytokines, eg, interferon-beta In viral clearance and persistence. However, knowledge of the role that RIG-I plays in HBV infection is limited. The woodchuck is a valuable model for studying HBV infection. To characterize the molecular basis of woodchuck RIG-I (w RIG-I) , we analyzed the complete coding sequences (CDSs) of w RIG-I, containing 2778 base pairs that encode 925 amino acids. The deduced w RIG-I protein was 106.847 k D with a theoretical isoelectric point (p I) of 6.07, and contained three important functional structures [caspase activation and recruitment domains (CARDs), DEx D / H-box helicases, and a repressor domain (RD)]. In woodchuck fibroblastoma cell line (WH12 / 6), w RIG-I-targeted small interfering RNA (si RNA) down-regulated RIG-I and its downstrean effector-IFN-beta transcripts under RIG-I ’ligand, 5’-ppp double stranded RNA (ds RNA) stimulation. We also measured m RNA levels of w RIG-I in different tissues from healthy woodchucks and in the livers from woodchuck hepatitis virus (WHV) -infected woodchucks . The basal expression levels of w RIG-I were abundant in the kidney and liver. Importantly, w RIG-I was significantly up-regulated in acutely infected woodchuck livers, suggesting that RIG-I might be involved in WHV infection. These results may characterize RIG-I in the woodchuck model, providing a strong basis for further study on RIG-I-mediated innate immunity in HBV infection.
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