Systematic profiling of protein complex dynamics reveals DNA-PK phosphorylation of IFI16 en route to herpesvirus immunity

• Published in: Science advances Vol. 7; no. 25
• Main Authors: Justice, Joshua L, Kennedy, Michelle A, Hutton, Josiah E, Liu, Dawei, Song, Bokai, Phelan, Brett, Cristea, Ileana M
• Format: Journal Article
• Language: English
• Published: United States American Association for the Advancement of Science 01-06-2021
• Subjects: Herpes Simplex; Herpesviridae; Herpesviridae Infections; Herpesvirus 1, Human; Host-Pathogen Interactions; Humans; Nuclear Proteins - genetics; Phosphoproteins; Phosphorylation; SciAdv r-articles; Systems Biology; Virology
• ISSN: 2375-2548; 2375-2548
• DOI: 10.1126/sciadv.abg6680

Dynamically shifting protein-protein interactions (PPIs) regulate cellular responses to viruses and the resulting immune signaling. Here, we use thermal proximity coaggregation (TPCA) mass spectrometry to characterize the on-off behavior of PPIs during infection with herpes simplex virus 1 (HSV-1), a virus with an ancient history of coevolution with hosts. Advancing the TPCA analysis to infer associations de novo, we build a time-resolved portrait of thousands of host-host, virus-host, and virus-virus PPIs. We demonstrate that, early in infection, the DNA sensor IFI16 recruits the active DNA damage response kinase, DNA-dependent protein kinase (DNA-PK), to incoming viral DNA at the nuclear periphery. We establish IFI16 T149 as a substrate of DNA-PK upon viral infection or DNA damage. This phosphorylation promotes IFI16-driven cytokine responses. Together, we characterize the global dynamics of PPIs during HSV-1 infection, uncovering the co-regulation of IFI16 and DNA-PK functions as a missing link in immunity to herpesvirus infection.