Molecular cloning, functional characterization of duck TRADD and its effect on infection with duck Tembusu virus

Tumor necrosis factor receptor 1 (TNFR1) associated death domain protein (TRADD) is a pivotal adaptor in TNF signaling pathway and plays an important role in apoptosis and immune regulation. The function of TRADD has been investigated extensively in mammals, however, the role of TRADD in ducks remains obscure. To reveal the function of duck TRADD (duTRADD) in the apoptosis and innate immune response, the TRADD homologue of mallard (Anas platyrhynchos) has been cloned and the function of duTRADD is investigated in this study. We conducted sequence analysis of the duTRADD, the open reading frame (ORF) region of duTRADD gene was 1065 bp, encoding 354 amino acids (aa), which shares similar functional domain to its mammalian counterpart. Tissue distribution profile of duTRADD in 7-day-old ducklings showed that the expression level of the gene was the highest in heart, followed by liver and brain. Accordingly, duck Tembusu virus (DTMUV) has been shown to decrease duTRADD expression, while overexpression of duTRADD inhibited DTMUV replication in a dose -dependent manner. Furthermore, duTRADD activated the transcriptional activity of caspase-3/8/9, the flow cytometry showed that duTRADD significantly induced apoptosis. However, duTRADD showed hardly any effect on the transcriptional activity of IFN-alpha/beta and its downstream interferon-stimulated genes (ISGs). The current data support the conclusion that duTRADD is a novel pro-apoptotic protein with a critical role in defense against DTMUV invasion. These results lay the theoretical foundation for the development of new anti-DTMUV strategies.