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Systematic profiling of protein complex dynamics reveals DNA-PK phosphorylation of IFI16 en route to herpesvirus
Joshua L Justice1, Michelle A Kennedy1, Josiah E Hutton1
1Department of Molecular Biology, Princeton University, Lewis Thomas Laboratory, Washington Road, Princeton, NJ 08544, USA.
Abstract:
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.
Insights
This study reveals how herpes simplex virus 1 (HSV-1) infection alters protein interactions. Researchers identified key protein dynamics, including the IFI16-DNA-PK interaction, crucial for immune response to viral DNA.
Area of Science:
- Immunology
- Virology
- Molecular Biology
Background:
- Protein-protein interactions (PPIs) are vital for cellular functions, including immune responses to viral infections.
- Herpes simplex virus 1 (HSV-1) has a long co-evolutionary history with hosts, suggesting complex interactions.
- Understanding dynamic PPIs during infection is crucial for deciphering host-pathogen interactions.
Purpose of the Study:
- To characterize the dynamic changes in PPIs during HSV-1 infection using a novel approach.
- To build a time-resolved map of host-host, virus-host, and virus-virus interactions.
- To elucidate the role of specific protein interactions in antiviral immunity.
Main Methods:
- Utilized thermal proximity coaggregation (TPCA) mass spectrometry to analyze PPIs.
- Developed advanced TPCA analysis to infer PPIs de novo.
- Investigated protein interactions during HSV-1 infection in a time-resolved manner.
Main Results:
- Generated a comprehensive, time-resolved map of thousands of PPIs during HSV-1 infection.
- Demonstrated that the DNA sensor IFI16 recruits DNA-dependent protein kinase (DNA-PK) to viral DNA early in infection.
- Identified IFI16 T149 as a substrate of DNA-PK, with phosphorylation promoting IFI16-mediated cytokine responses.
Conclusions:
- The dynamic interplay of PPIs is central to cellular responses against HSV-1.
- The co-regulation of IFI16 and DNA-PK is a critical, previously unrecognized component of anti-herpesvirus immunity.
- This research provides a new framework for understanding viral infection dynamics at the molecular level.
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