Molecular characterization and expression of TAK-binding proteins (TAB1-3) in Larimichthys crocea infected by Vibrio parahemolyticus and LPS

Abstract

TAK1-binding proteins (TABs) are important immune protein involved in various intracellular signalling pathways. Here, TAB1-3 (lcTAB1-3) were characterized from Larimichthys crocea. The predicted 1524 bp coding sequence of lcTAB1 encoded a 507-residue protein, while lcTAB2 (2271 bp) and lcTAB3 (1836 bp) encoded 756 and 611 residue proteins, respectively. Their sequence shared conserved domain structures and functional sites with their orthologs from other species. The expression of lcTAB1-3 were detected in all tested tissues, which were upregulated in spleen, liver and kidney following Vibrio parahemolyticus infection. Immunofluorescence staining revealed that lcTAB1 were localized in cytoplasm, while lcTAB2 and lcTAB3 were in the endsome. Moreover, the NF-κB protein level was obviously upregulated after the co-overexpression of lcTAK1 and lcTABs, higher than that after the overexpression of lcTAK1 or lcTABs alone. Co-immunoprecipitation proved the direct interaction of lcTAB1/lcTAB2/lcTAB3 and lcTAK1. These findings indicated the roles of lcTABs in immune response of Larimichthys crocea.

Keywords: Co-immunoprecipitation; Immunofluorescence staining; Larimichthys crocea; TAK1-Binding proteins; Western blot; qRT-PCR.