African swine fever virus pMGF505-9R enhances RIG-I-like receptor signaling by promoting RING finger protein 125 autoubiquitination

African swine fever (ASF) is a highly infectious disease that poses a significant threat to the global pig industry. Recent studies have demonstrated that the African swine fever virus (ASFV) infection can cause severe inflammatory responses and promote the production of cytokines, but it is still unclear whether the viral proteins play a role in this process. Therefore, we conducted a genome-wide screening by dual luciferase activity assay. The results showed that six viral proteins of ASFV have a stimulating effect on the promoter activity of interferon beta (IFN-β). Among them, the nonstructural protein pMGF505-9R significantly increased the activity of IFN-β promoter and promoted the expression of IFN-β. In addition, pMGF505-9R can also increase the promoter activity of NF-κB and promoted the expression of interleukin 1 beta (IL-1β). Further research found that pMGF505-9R could regulate the retinoic acid-inducible gene I-like receptor signaling pathways by influencing the protein level of RING finger protein 125 (RNF125), ultimately affecting the expression of IFN-β and IL-1β. Mechanistically, pMGF505-9R interacted with the host E3 ubiquitin ligase RNF125. This interaction promoted the autoubiquitination and subsequent degradation of RNF125, which in turn reduced the K48 ubiquitination of retinoic acid-inducible gene I, melanoma differentiation-associated protein 5, and mitochondrial antiviral signaling, which ultimately promoted the production of IFN-β and IL-1β. The results provide novel insights into the immune regulatory mechanisms of ASFV, which would greatly improve our understanding of virus pathogenesis.