Workflows and considerations for investigating protein interactions of viral DNA sensors

Timothy R. Howard, Bokai Song, Ileana M. Cristea

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

DNA sensors are a core component of innate immunity in mammalian cells. In response to pathogen infection, these specialized proteins sense pathogenic DNA from bacteria or viruses and initiate immune signaling cascades. These defense mechanisms rely on the rapid formation and temporal regulation of protein-protein interactions. Similarly, protein interactions underlie virus immune evasion mechanisms, as proteins from diverse viruses associate with and inhibit DNA sensors. Here, we describe experimental protocols for identifying protein interactions of DNA sensors, and discuss considerations for optimal isolation of protein complexes when targeting either endogenous or tagged proteins. Additionally, as viral infections and immune responses are known to induce prominent changes in cellular protein abundances, we provide a workflow for investigating these protein associations in the context of proteome alterations.

Original languageEnglish (US)
Title of host publicationMethods in Enzymology
EditorsJungsan Sohn
PublisherAcademic Press Inc.
Pages309-338
Number of pages30
ISBN (Print)9780128183595
DOIs
StatePublished - 2019

Publication series

NameMethods in Enzymology
Volume625
ISSN (Print)0076-6879
ISSN (Electronic)1557-7988

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

Keywords

  • DNA sensing
  • DNA virus
  • IFI16
  • IP
  • Immunoaffinity purification
  • Mass spectrometry
  • Protein interactions
  • Proteome
  • Proteomics
  • cGAS

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  • Cite this

    Howard, T. R., Song, B., & Cristea, I. M. (2019). Workflows and considerations for investigating protein interactions of viral DNA sensors. In J. Sohn (Ed.), Methods in Enzymology (pp. 309-338). (Methods in Enzymology; Vol. 625). Academic Press Inc.. https://doi.org/10.1016/bs.mie.2019.04.026