P300/RNA polymerase II mediates induction of the teleost viral RNA sensor MDA5 through the interferon regulatory factor IRF11.

Melanoma differentiation-associated gene 5 (MDA5) initiates type I interferon (IFN) production by detecting cytosolic viral RNA. Mammalian MDA5 is an IFN-inducible gene and controlled by IFN regulatory factor 1 (IRF1). Teleost MDA5 also induces type I IFN production in response to viruses, yet its regulation remains largely unexplored. This study used the large yellow croaker Larimichthys crocea (Lc) as a model organism and revealed that a type I IFN (LcIFNi) triggers the expression of LcMDA5 through the JAK-STAT signaling pathway, which involves phosphorylation of LcIRF11. LcMDA5 was transcriptionally regulated by LcIRF11. Mechanistically, LcIRF11 interacts with the IFN-stimulated response element (ISRE) within the LcMDA5 promoter, via α3 helix and loop1, and loop2 and loop3 in its DNA binding domain (DBD). Overexpression of LcIRF11 recruits p300 and RNA polymerase II (Pol II) to the LcMDA5 promoter region. Pull-down analysis further confirmed the interaction of LcIRF11 with these two proteins. This recruitment was accompanied by increased levels of histone H3K27 acetylation (H3K27ac) and histone H3K4 trimethylation (H3K4me3), both of which are strongly associated with active transcription. Conversely, silencing LcIRF11 reduced p300 and Pol II recruitments and hindered the enrichment of H3K27ac/H3K4me3 modifications at the LcMDA5 promoter. Thus, here we present the first report of IRF11 orchestrating the activation of MDA5 transcription by binding to the ISRE of MDA5 promoter and forming a transcriptional complex with p300 and Pol II. Our results revealed an ancient regulatory mechanism of MDA5 in lower vertebrates, providing insights into its function and evolution.