Workflows and considerations for investigating protein interactions of viral DNA sensors.

TitleWorkflows and considerations for investigating protein interactions of viral DNA sensors.
Publication TypeJournal Article
Year of Publication2019
AuthorsHoward, TR, Song, B, Cristea, IM
JournalMethods Enzymol
Volume625
Pagination309-338
Date Published2019
ISSN1557-7988
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.

DOI10.1016/bs.mie.2019.04.026
Alternate JournalMeth. Enzymol.
PubMed ID31455534
PubMed Central IDPMC6889811
Grant ListR01 GM114141 / GM / NIGMS NIH HHS / United States
T32 GM007388 / GM / NIGMS NIH HHS / United States