Molecular characterization and expression analyses of two homologues of interferon-stimulated gene ISG15 in Larimichthys crocea (Family: Sciaenidae).

In this study, we sequenced and characterized two homologues of interferon-stimulated gene ISG15, termed as LcISG15-1 and LcISG15-2, from the large yellow croaker (Larimichthys crocea). The LcISG15-1 encodes 159 amino acids and the LcISG15-2 encodes 155 amino acids, both of which contain two tandem ubiquitin-like domains and the conserved C-terminal LRGG conjugation motif. The sequence analyses showed that both the LcISG15-1 and LcISG15-2 exhibit high similarity with ISG15 from other fishes. A putative IFN-stimulatory response element (ISRE) was detected in promoter regions of both the LcISG15-1 and LcISG15-2. Phylogenetic analyses revealed a close evolutionary relationship of both the LcISG15-1 and LcISG15-2 with other teleostean ISG15. Molecular evolutionary analyses suggested a gene duplication event of ISG15 in the ancestor of the Sciaenidae, with a signature of positive selection was found in the ISG15-2 gene copy of sciaenid fishes. The Real-time PCR analyses showed that the LcISG15-1 and LcISG15-2 were both found to be ubiquitously expressed in ten examined organs in large yellow croaker, with predominant expressions both in peripheral blood. Expression analyses showed that both the LcISG15-1 and LcISG15-2 were rapidly and significantly upregulated in vivo after poly (I:C) challenge in liver and spleen organs. However, the LcISG15-1 and LcISG15-2 were both significantly induced after pathogen Vibrio parahemolyticus infection only in the liver but not in the spleen. These results indicated that there are two ISG15 homologues in the large yellow croaker, both of which are likely to be involved in host immune defense against viral and bacterial infection.