Cloning, expression, and bioinformatics analysis of a putative pigeon retinoid acid-inducible gene-I

Abstract

Retinoid acid-inducible gene-I (RIG-I) is a major cytoplasmic RNA sensor, playing an essential role in detecting viral RNA and triggering antiviral innate immune responses. The objective of the present study was to characterize the structure and expression of the RIG-I gene in pigeons. The pigeon RIG-I (piRIG-I) was cloned from splenic lymphocytes of pigeons using reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends. The cDNA of piRIG-I contains a 147 bp 5′-untranslated regions (UTRs), a 2787 bp open reading frame, and 2962 bp 3′-UTRs. Based on this sequence, the encoded piRIG-I protein is predicted to consist of 928 amino acids, and it has conserved domains typical of RIG-I-like receptors (RLRs) including two tandem arranged N-terminal caspase recruitment domains, a domain with the signature of DExD/H box helicase (helicase domain), and a C-terminal repression domain similar to finch RIG-I, duck RIG-I, goose RIG-I, human RIG-I, and mouse RIG-I. The piRIG-I shows 82.1%, 78.6%, and 78.2% amino acid sequence identity with previously described finch RIG-I, duck RIG-I, and goose RIG-I, respectively, and 49.7%–53.8% sequence identity with mammalian homologs. Quantitative RT-PCR (qRT-PCR) analysis indicated that the piRIG-I mRNA is scarcely detected in healthy tissues, and it is strongly expressed in the thymus gland, kidney, spleen, and bursa of Fabricius. These findings lay the foundation for further research on the function and mechanism of avian RIG-I in innate immune response related to vaccinations and infectious diseases in the pigeon.