Molecular characterization, tissue distribution and functional analysis of galectin 1-like 2 in grass carp (Ctenopharyngodon idella).

ABSTRACT

Galectins, as an evolutionary conserved group of lectin superfamily, has the functions of pathogen recognition, anti-bacteria and anti-virus. In this study, a 405 bp cDNA sequence of galectin 1-like 2 (CiGal1-L2) was obtained from grass carp (Ctenopharyngodon idella), which encoded 134 amino acids with a predicted molecular mass of 15.143 kDa and an isoelectric point of 5.33. The sugar binding motifs (H-N-R, V-N and W--E-R) were detected in carbohydrate-binding domain (CRD). The amino acid sequence similarity showed that CiGal1-L2 was 40.30-42.54% and 66.42-81.20% similarity to mammalian and fish counterparts, respectively. The phylogenetic tree showed that CiGal1-L2 was clustered with fish galectin-1s and closely related to Cyprinus carpio. Real-time quantitative PCR (RT-qPCR) analysis revealed that CiGal1-L2 was widely expressed in all tested tissues. In addition, the expression of CiGal1-L2 was differentially up-regulated challenged with grass carp reovirus (GCRV), lipopolysaccharide (LPS) and polyinosinicpolycytidylic acid (poly IC). The fluorescence of CiGal1-L2-GFP was distributed in the cytoplasm and nucleus of HEK 293T cells and showed a trend of nuclear translocation after LPS and poly IC treatment. Finally, the recombinant CiGal1-L2 (rCiGal1-L2) protein showed strong binding ability to LPS. In conclusion, the results provided further insight into the immune roles of galectin-1 in teleost.