Abstract

As a member of DExD/H-box helicase family, DDX41 (DEAD box polypeptide 41) acts as an intracellular DNA sensor that induces type I IFN expression in mammals. Fish DDX41 shares some similar properties with the mammalian counterparts. In this study, a DDX41 orthologous gene from grass carp (Ctenopharyngodon idellus) (CiDDX41) was cloned and characterized. The ORF of CiDDX41 encodes a polypeptide of 614 amino acids. Multiple alignments showed that DDX41 is highly conserved among different species. Phylogenetic tree analysis revealed that CiDDX41 shares a high degree of homology with Sinocyclocheilus rhinocerous DDX41. CiDDX41 is highly expressed in kidney, intestines, liver and spleen. Their expressions are up-regulated more obviously after the treatment with GCRV. Over-expression of CiDDX41 in CIK cells increases the transcription level of grass carp IFN I and ISG15. On the contrary, knockdown of CiDDX41 inhibits the IFN I and ISG15 transcription. Moreover, a part of CiDDX41 translocates from the nuclear to cytoplasm to interact with grass carp STING (CiSTING). In CIK cells, overexpression of CiDDX41 and CiSTING can promote the phosphorylation and nuclear-cytoplasm translocation of grass carp IRF7 (CiIRF7) and then acutely up-regulate the IFN I and ISG15 expression. However, the knockdown of CiDDX41 inhibits the phosphorylation IRF7. Taken together, all these results above suggested that CiDDX41 performs as an activator for innate immune through STING-IRF7 mediated signaling pathway.

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