Skip to main content

Discovery of Host–Viral Protein Complexes During Infection

Author(s): Rowles, Daniell L; Terhune, Scott S; Cristea, Ileana M.

To refer to this page use:
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.
Publication Date: 1-Jan-2013
DOI: doi:10.1007/978-1-62703-601-6_4
EISSN: 1940-6029
Keywords: virus-host interactions, infection, epitope tag, immunoaffinity purification, cryogenic cell lysis, proteomics, protein complex, protein-protein interactions, mass spectrometry
Language: eng
Type of Material: Journal Article
Journal/Proceeding Title: Methods in Molecular Biology
Version: Author's manuscript

Items in OAR@Princeton are protected by copyright, with all rights reserved, unless otherwise indicated.