The categorization and mutual modulation of expanded MyD88s in Crassostrea gigas

•MyD88s expanded in Crassostrea gigas were categorized according to their sequence and functional characters.•CgMyD88s in same type shared similar spatio-temporal expression profiles.•Type I CgMyD88 positively contributed to the activation of NF-B signaling.•Type II and Type III CgMyD88s suppressed the activity of Type I on the activation of NF-B signaling.

MyD88 serves as a critical cytosolic adaptor mediating activation of NF-κB in innate immunity. It has been found that there is a considerable expansion of MyD88 in Crassostrea gigas. In the present study, four typical MyD88 genes in Crassostrea gigas (CgMyD88-A to CgMyD88-D) were successfully cloned and their potential functions were investigated together with another two known ones (CgMyD88-T1 and CgMyD88-T2). Multiple alignments revealed that CgMyD88-B and CgMyD88-C remained the conserved DD and TIR domains, while there was a significant variation of E51Q in the DD of CgMyD88-A, and some variations in both DD and TIR domains of CgMyD88-D, respectively. Both truncated CgMyD88-T1 and CgMyD88-T2 lacked Box II in their only TIR domains. Expression pattern analysis showed that CgMyD88-B and CgMyD88-C genes possessed higher expression in normal tissues, compared with the other four. When oysters were under bacteria challenge, CgMyD88-B, CgMyD88-C, CgMyD88-T1 and CgMyD88-T2 were firstly induced, while CgMyD88-A and CgMyD88-D were suppressed. Dual luciferase reporter assays showed that CgMyD88-B and CgMyD88-C could promote the activation of NF-κB signaling pathway, while the other four CgMyD88 genes failed or even suppressed the activities of CgMyD88-B and CgMyD88-C on the activation of NF-κB signaling. It was deduced that after oysters were challenged by bacteria, CgMyD88-B and CgMyD88-C could rapidly and efficiently activate NF-κB signaling pathway to elicit anti-pathogen responses before suppressor CgMyD88 genes (CgMyD88-T1 and CgMyD88-T2) exceeding their expression level. These results suggested that there was mutual modulation of expanded CgMyD88 genes on activating NF-κB signaling pathway in oyster C. gigas.