title: Resolution of the cellular proteome of the nucleocapsid protein from a highly pathogenic isolate of porcine reproductive and respiratory syndrome virus identifies PARP-1 as a cellular target whose interaction is critical for virus biology.
abstract: Porcine reproductive and respiratory syndrome virus (PRRSV) is a major threat to the swine industry and food security worldwide. The nucleocapsid (N) protein is a major structural protein of PRRSV. The primary function of this protein is to encapsidate the viral RNA genome, and it is also thought to participate in the modulation of host cell biology and recruitment of cellular factors to facilitate virus infection. In order to the better understand these latter roles the cellular interactome of PRRSV N protein was defined using label free quantitative proteomics. This identified several cellular factors that could interact with the N protein including poly [ADP-ribose] polymerase 1 (PARP-1), a cellular protein, which can add adenosine diphosphate ribose to a protein. Use of the PARP-1 small molecule inhibitor, 3-AB, in PRRSV infected cells demonstrated that PARP-1 was required and acted as an enhancer factor for virus biology. Serial growth of PRRSV in different concentrations of 3-AB did not yield viruses that were able to grow with wild type kinetics, suggesting that by targeting a cellular protein crucial for virus biology, resistant phenotypes did not emerge. This study provides further evidence that cellular proteins, which are critical for virus biology, can also be targeted to ablate virus growth and provide a high barrier for the emergence of drug resistance.
authors.affiliation: College of Life Sciences, Northwest A&F University, Yangling, China.
authors.forename: Long
authors.initials: L
authors.lastname: Liu
authors.affiliation: School of Molecular and Cellular Biology and Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds, UK.
authors.forename: Zoe
authors.initials: Z
authors.lastname: Lear
authors.affiliation: School of Molecular and Cellular Biology and Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds, UK.
authors.forename: David J
authors.initials: DJ
authors.lastname: Hughes
authors.affiliation: Department of Infection Biology, Institute of Infection and Global Health, University of Liverpool, Liverpool, UK.
authors.forename: Weining
authors.initials: W
authors.lastname: Wu
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, Yangling, China.
authors.forename: En-min
authors.initials: EM
authors.lastname: Zhou
authors.affiliation: School of Molecular and Cellular Biology and Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds, UK.
authors.forename: Adrian
authors.initials: A
authors.lastname: Whitehouse
authors.affiliation: College of Life Sciences, Northwest A&F University, Yangling, China. Electronic address: chenhy@nwsuaf.edu.cn.
authors.forename: Hongying
authors.initials: H
authors.lastname: Chen
authors.affiliation: Department of Infection Biology, Institute of Infection and Global Health, University of Liverpool, Liverpool, UK; College of Veterinary Medicine, Northwest A&F University, Yangling, China; NIHR Health Protection Research Unit in Emerging and Zoonotic Infections, Liverpool, UK. Electronic address: julian.hiscox@liverpool.ac.uk.
authors.forename: Julian A
authors.initials: JA
authors.lastname: Hiscox
mesh_terms: D000818:Animals; D005786:Gene Expression Regulation; D019590:Nucleocapsid Proteins / Q000235:genetics / Q000378:metabolism*; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism*; D019316:Porcine respiratory and reproductive syndrome virus / Q000235:genetics / Q000378:metabolism* / Q000472:pathogenicity; D011485:Protein Binding; D020543:Proteome / Q000378:metabolism*; D040901:Proteomics; D013552:Swine; D014774:Virulence
chemical_list: D019590:Nucleocapsid Proteins; D020543:Proteome; D011065:Poly(ADP-ribose) Polymerases
keywords: 3-AB; Antivirus; Nucleocapsid; PARP-1; PRRSV; Proteome