title: Discovery of host-viral protein complexes during infection.
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.
authors.affiliation: Department of Molecular Biology, Princeton University, Princeton, NJ, USA.
authors.forename: Daniell L
authors.initials: DL
authors.lastname: Rowles
authors.forename: Scott S
authors.initials: SS
authors.lastname: Terhune
authors.forename: Ileana M
authors.initials: IM
authors.lastname: Cristea
mesh_terms: D000818:Animals; D002846:Chromatography, Affinity / Q000379:methods; D054884:Host-Pathogen Interactions; D006801:Humans; D007239:Infections / Q000378:metabolism* / Q000821:virology*; D046912:Multiprotein Complexes / Q000302:isolation & purification / Q000378:metabolism*; D025941:Protein Interaction Mapping / Q000379:methods*; D040901:Proteomics / Q000379:methods; D014764:Viral Proteins / Q000378:metabolism*; D014780:Viruses / Q000378:metabolism*
chemical_list: D046912:Multiprotein Complexes; D014764:Viral Proteins