Fas-associated death domain (FADD) in sea cucumber (Apostichopus japonicus): Molecular cloning, characterization and pro-apoptotic function analysis

Abstract

Fas-associated death domain (FADD) is an adaptor protein that functions in transferring the apoptotic signals regulated by the death receptors. In this study, a full-length cDNA of FADD homologue in sea cucumber Apostichopus japonicas (AjFADD) was cloned and characterized, and its functional roles in apoptosis investigated. In healthy sea cucumbers, AjFADD was expressed in all detected tissues, with higher levels in coelomocytes and intestine. AjFADD mRNA and protein levels were significantly expressed in coelomocytes after exposed with LPS or poly (I:C) in vitro, and challenged with Vibrio splendidus in vivo. Moreover, siRNA-mediated AjFADD knockdown in coelomocyte much decreased AjFADD mRNA and protein levels as well as the coelomocytes apoptosis levels. Furthermore, over-expression of the expression plasmid pcDNA3.1 encoding AjFADD (pcAjFADD) significantly increased the apoptosis levels in HEK293 cells. Taken together, our results support that AjFADD is a novel pro-apoptotic protein that might play key roles in defensing the bacterial and virus invasion in sea cucumber.