Discovery of host-viral protein complexes during infection

Daniell L. Rowles, Scott S. Terhune, Ileana M. Cristea

Research output: Chapter in Book/Report/Conference proceedingChapter

13 Scopus citations

Abstract

Viruses have co-evolved with their hosts, developing effective approaches for hijacking and manipulating host cellular processes. Therefore, for their efficient replication and spread, viruses depend on dynamic and temporally regulated interactions with host proteins. The rapid identification of host proteins targeted by viral proteins during infection provides significant insights into mechanisms of viral protein function. The resulting discoveries often lead to unique and innovative hypotheses on viral protein function. Here, we describe a robust method for identifying virus-host protein interactions and protein complexes, which we have successfully utilized to characterize spatial-temporal protein interactions during infections with either DNA or RNA viruses, including human cytomegalovirus (HCMV), herpes simplex virus type 1 (HSV-1), pseudorabies virus (PRV), human immunodeficiency virus (HIV-1), Sindbis, and West Nile virus (WNV). This approach involves cryogenic cell lysis, rapid immunoaffinity purification targeting a virus or host protein, followed by identification of associated proteins using mass spectrometry. Like most proteomic approaches, this methodology has evolved over the past few years and continues to evolve. We are presenting here the updated approaches for each step, and discuss alternative strategies allowing for the protocol to be optimized for different biological systems.

Original languageEnglish (US)
Title of host publicationVirus-Host Interactions
Subtitle of host publicationMethods and Protocols
EditorsSusanne Bailer, Diana Lieber
Pages43-70
Number of pages28
DOIs
StatePublished - 2013

Publication series

NameMethods in Molecular Biology
Volume1064
ISSN (Print)1064-3745

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

Keywords

  • Cryogenic cell lysis
  • Epitope tag
  • Immunoaffinity purification
  • Infection
  • Mass spectrometry
  • Protein complex
  • Protein-protein interactions
  • Proteomics
  • Virus-host interactions

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