Molecular characterization, expression patterns, and subcellular localization of RIG-I in the Jinding duck (Anas platyrhynchos domesticus)

Abstract

Retinoic acid-inducible gene I (RIG-I)-like receptors (RLRs) have recently been identified as cytoplasmic sensors for RNA virus. Recent research has shown that RIG-I, a member of this family, play an important role in innate immunity. In this study, we cloned the RIG-I gene from Jinding duck by reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE). We determined that the cDNA of duRIG-I contains a 14-bp 5' UTR, a 2802-bp open reading frame, and alternative 3' UTRs (295-bp and 927-bp) and encodes a polypeptide of 933 amino acids. Based on this sequence, the duRIG-I protein is predicted to have conserved domains typical of RLRs. In addition, duRIG-I was found to be distributed throughout DF1 cells by indirect immunofluorescence, as predicted. duRIG-I mRNA was scarcely detected in healthy tissues by semi-quantitative RT-PCR (sqRT-PCR). To study the role of RIG-I in innate immunity, we used synthetic double-stranded RNA to mimic viral infection in vivo and detected duRIG-I transcripts in spleen and liver by quantitative real-time PCR (qRT-PCR). The expression of duRIG-I mRNA was significantly elevated at 8h post-injection (P < 0.05) and was indistinguishable from control levels at other time points (P > 0.05). These results suggest that duRIG-I plays an important role in innate immune responses to double-stranded RNA viruses and warrant further studies to reveal the possible mechanism.

Keywords: Duck; Expression pattern; RIG-I; Subcellular localization.