DNA-PK senses DNA virus infection in human cells

Abstract The type I interferon (IFN-I) response to virus infection is initiated by the detection of nucleic acids by intracellular pattern recognition receptors (PRRs). The sensing of DNA viruses by PRRs is carried out by a number of DNA-binding PRRs that signal via the adaptor protein stimulator of interferon genes (STING) to drive IFN-I transcription. We previously described the role of DNA-dependent protein kinase (DNA-PK) as a viral DNA sensor in murine fibroblasts. In this study we show that DNA-PK is essential for the host response to DNA and DNA viruses in human fibroblasts. In the absensce of the catalytic subunit of the DNA-PK heterotrimer, DNA-PKcs, fibroblasts are deficient in their ability to activate a IFN-I response to DNA. DNA-PKcs is activated rapidly following exogenous DNA transfection or DNA virus infection and is required for signalling via STING and the kinase TBK-1 to the transcription factor interferon regulator factor 3 (IRF-3). Most wild-type DNA viruses combat intracellular DNA PRRs using immunomodulatory proteins encoded in their genomes and are effective at blocking IFN-I responses in infected cells. Here we make use of attenutated vaccina and herpes simplex 1 viruses that are lacking the immunomodulators that target DNA sesining mechanisms. We show that DNA-PKcs can sense these viruses and is required for triggering the IFN-I response in infected cells. These data cement the role of DNA-PK in the sensing of DNA virus infections in human cells.