title: Characterization of the Interactome of the Porcine Reproductive and Respiratory Syndrome Virus Nonstructural Protein 2 Reveals the Hyper Variable Region as a Binding Platform for Association with 14-3-3 Proteins.
abstract: Porcine reproductive and respiratory syndrome virus (PRRSV) is a major threat to the swine industry worldwide and hence global food security, exacerbated by a newly emerged highly pathogenic (HP-PRRSV) strain from China. PRRSV nonstructural protein 2 (nsp2) is a multifunctional polypeptide with strain-dependent influences on pathogenicity. A number of discrete functional regions have been identified on the protein. Quantitative label free proteomics was used to identify cellular binding partners of nsp2 expressed by HP-PRRSV. This allowed the identification of potential cellular interacting partners and the discrimination of nonspecific interactions. The interactome data were further investigated and validated using biological replicates and also compared with nsp2 from a low pathogenic (LP) strain of PRRSV. Validation included both forward and reverse pulldowns and confocal microscopy. The data indicated that nsp2 interacted with a number of cellular proteins including 14-3-3, CD2AP, and other components of cellular aggresomes. The hyper-variable region of nsp2 protein was identified as a binding platform for association with 14-3-3 proteins.
authors.affiliation: Department of Infection Biology, Institute of Infection and Global Health, University of Liverpool , Liverpool L3 5RF, U.K.|Department of Basic Veterinary Medicine, College of Animal Science and Veterinary Medicine, Shandong Agricultural University , Tai'an 271018, P.R. China.
authors.forename: Yihong
authors.initials: Y
authors.lastname: Xiao
authors.affiliation: Department of Infection Biology, Institute of Infection and Global Health, University of Liverpool , Liverpool L3 5RF, U.K.
authors.forename: Weining
authors.initials: W
authors.lastname: Wu
authors.affiliation: College of Veterinary Medicine, Northwest A&F University , Yangling 712100, P.R. China.
authors.forename: Jiming
authors.initials: J
authors.lastname: Gao
authors.affiliation: The Roslin Institute, University of Edinburgh , Edinburgh EH25 9RG, U.K.
authors.forename: Nikki
authors.initials: N
authors.lastname: Smith
authors.affiliation: The Roslin Institute, University of Edinburgh , Edinburgh EH25 9RG, U.K.
authors.forename: Christine
authors.initials: C
authors.lastname: Burkard
authors.affiliation: Department of Infection Biology, Institute of Infection and Global Health, University of Liverpool , Liverpool L3 5RF, U.K.
authors.forename: Dong
authors.initials: D
authors.lastname: Xia
authors.affiliation: Department of Basic Veterinary Medicine, College of Animal Science and Veterinary Medicine, Shandong Agricultural University , Tai'an 271018, P.R. China.
authors.forename: Minxia
authors.initials: M
authors.lastname: Zhang
authors.affiliation: College of Veterinary Medicine, Northwest A&F University , Yangling 712100, P.R. China.
authors.forename: Chengbao
authors.initials: C
authors.lastname: Wang
authors.affiliation: The Roslin Institute, University of Edinburgh , Edinburgh EH25 9RG, U.K.
authors.forename: Alan
authors.initials: A
authors.lastname: Archibald
authors.affiliation: The Roslin Institute, University of Edinburgh , Edinburgh EH25 9RG, U.K.
authors.forename: Paul
authors.initials: P
authors.lastname: Digard
authors.affiliation: College of Veterinary Medicine, Northwest A&F University , Yangling 712100, P.R. China.
authors.forename: En-Min
authors.initials: EM
authors.lastname: Zhou
authors.affiliation: Department of Infection Biology, Institute of Infection and Global Health, University of Liverpool , Liverpool L3 5RF, U.K.|College of Veterinary Medicine, Northwest A&F University , Yangling 712100, P.R. China.
authors.forename: Julian A
authors.initials: JA
authors.lastname: Hiscox
mesh_terms: D048948:14-3-3 Proteins / Q000378:metabolism*; D000818:Animals; D001665:Binding Sites; D002460:Cell Line; D054884:Host-Pathogen Interactions; D006801:Humans; D019316:Porcine respiratory and reproductive syndrome virus / Q000737:chemistry* / Q000472:pathogenicity; D025941:Protein Interaction Mapping; D013552:Swine; D017361:Viral Nonstructural Proteins / Q000378:metabolism*
chemical_list: D048948:14-3-3 Proteins; D017361:Viral Nonstructural Proteins
keywords: aggresomes; interactome; label-free proteomics; nonstructural protein 2; porcine reproductive and respiratory syndrome virus; proteomics; virus