PT - JOURNAL ARTICLE AU - Katelyn Burleigh AU - Joanna H. Maltbaek AU - Stephanie Cambier AU - Richard Green AU - Michael Gale, Jr. AU - Richard C. James AU - Daniel B. Stetson TI - Human DNA-PK activates a STING-independent DNA sensing pathway AID - 10.1101/587501 DP - 2019 Jan 01 TA - bioRxiv PG - 587501 4099 - http://biorxiv.org/content/early/2019/03/23/587501.short 4100 - http://biorxiv.org/content/early/2019/03/23/587501.full AB - Detection of intracellular DNA by the cGAS-STING pathway activates a type I interferon-mediated innate immune response that protects from virus infection and can be harnessed to promote anti-tumor immunity. Whether there are additional DNA sensing pathways, and how such pathways might function, remains controversial. We show here that humans – but not mice – have a second, potent, STING-independent DNA sensing pathway that is blocked by the E1A viral oncogene of human adenovirus 5. We identify human DNA-PK as the sensor of this pathway and demonstrate that DNA-PK kinase activity drives a robust and broad antiviral response. We discover that the heat shock protein HSPA8/HSC70 is a unique target of DNA-PK. Finally, we demonstrate that detection of foreign DNA and DNA damage trigger distinct modalities of DNA-PK activity. These findings reveal the existence, sensor, unique target, and viral antagonists of a STING-independent DNA sensing pathway (SIDSP) in human cells.