Molecular characterization and functional analysis of MyD88 from the tropical sea cucumber, Holothuria leucospilota

In this study, a myeloid differentiation factor 88 (MyD88) named as HLMyD88 was identified from the sea cucumber Holothuria leucospilota. The full-length cDNA of HLMyD88 is 4797 bp in size, containing a 227 bp 5'-untranslated region (UTR), a 3721 bp 3'-UTR and an 849 bp open reading frame (ORF) encoding a protein of 282 amino acids with a deduced molecular weight of 32.25 kDa HLMyD88 contains an N-terminal death domain and a C-terminal Toll/interluekin-1 receptor (TIR) domain with three highly conserved sequence motifs named as Box 1, Box 2 and Box 3. The results of luciferase reporter assay showed that over-expressed HLMyD88 in HEK293T cells could activate the transcription factors nuclear factor-kappa B (NF-kappa B) and activator protein 1 (AP-1). Additionally, the secretion of proinflammatory cytokines IL-1 beta and TNF-alpha in the HEK293T cells was increased by over-expressed HLMyD88, indicating the potential role of HLMyD88 in the innate immunity of sea cucumber. Moreover, we further confirmed that over-expressed HLMyD88 could also induce apoptosis.