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Interferon regulatory factors (IRFs) are transcription factors found in both vertebrates and invertebrates that were recently identified and found to play an important role in antiviral immunity in black tiger shrimp  Penaeus monodon . In this study, we investigated the mechanism by which  P. monodon  IRF ( Pm IRF) regulates the immune-related genes downstream of the cytosolic DNA sensing pathway. Depletion of  Pm IRF by double-stranded RNA-mediated gene silencing significantly reduced the mRNA expression levels of the IFN-like factors  Pm Vago1,  Pm Vago4, and  Pm Vago5 and antilipopolysaccharide factor 6 (ALF Pm 6 )  in shrimp. In human embryonic kidney (HEK293T) cells transfected with  Pm IRF or co-transfected with DEAD-box polypeptide ( Pm DDX41) and simulator of IFN genes ( Pm STING) expression plasmids, the promoter activity of IFN-β, nuclear factor (NF-κB), and ALF Pm 6 was synergistically enhanced following stimulation with the nucleic acid mimics deoxyadenylic–deoxythymidylic acid sodium salt [poly(dA:dT)] and high molecular weight (HMW) polyinosinic–polycytidylic acid [poly(I:C)]. Both nucleic acid mimics also significantly induced  Pm STING,  Pm IRF, and ALF Pm 6 gene expression. Co-immunoprecipitation experiments showed that  Pm IRF interacted with  Pm STING in cells stimulated with poly(dA:dT).  Pm STING,  Pm IRF, and  Pm DDX41 were localized in the cytoplasm of unstimulated HEK293T cells and  Pm IRF and  Pm DDX41 were translocated to the nucleus upon stimulation with the nucleic acid mimics while  Pm STING remained in the cytoplasm. These results indicate that  Pm IRF transduces the pathogen signal  via  the  Pm DDX41– Pm STING DNA sensing pathway to induce downstream production of interferon-like molecules and antimicrobial peptides.

Penaeus monodon Interferon Regulatory Factor (PmIRF) Activates IFNs and Antimicrobial Peptide Expression via a STING-Dependent DNA Sensing Pathway