Characterization of the interactome of the porcine reproductive and respiratory syndrome virus glycoprotein-5
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Porcine reproductive and respiratory syndrome virus (PRRSV) is one of the most important pathogens in the swine industry, causing reproductive failure in sows and respiratory disorders in piglets. Glycosylated protein 5 (GP5) is a major envelope protein of the virus. It is essential for virus particle assembly and involved in viral pathogenesis. In the present study, we identified the host cellular proteins that interact with GP5 by performing immunoprecipitation in MARC-145 cells infected by a recombinant PRRSV containing a FLAG-tag insertion in GP5. In total, 122 cellular proteins were identified by LC-MS/MS. Gene Ontology and KEGG databases were used to map these proteins to different cellular processes, locations and functions. Interestingly, 10.24% of identified cellular proteins were involved in the process of translation. Follow up experiments demonstrated that expression of GP5 in transfected cells led to inhibition of translation of reporter genes. Interaction between GP5 and ATP synthase subunit alpha (ATP5A) was further confirmed by co-immunoprecipitation suggesting a possible role of GP5 in regulation of ATP production in cells. These data contribute to a better understanding of GP5’s role in viral pathogenesis and virus-host interactions.
We thank Dr. X-J Meng (Virginia Tech) for supplying us with plasmid pIR-VR2385-CA. Also, we are thankful to Dr. Aviad Levin (University of Alberta) for designing a peptide for rabbit immunization, and Dr. Samuel Attah-Poku for making peptide conjugates. We acknowledge the Proteomics Center at the University of Missouri-Columbia for mass spectrometry analysis, and Dr. Aaron White for critical reading of the manuscript. This paper was published with the permission of the Director of VIDO-InterVac, journal series no. 817.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest. The study was funded in part by funding from the University of Saskatchewan President’s NSERC fund grant (A.Z. is a principal investigator), and M.Z. is supported by a China Scholarship Council.
Research involving human participants and/or animals
This article does not contain any studies with human participants. Work with animals was approved by the University of Saskatchewan’s Animal Research Ethics Board and adhered to the Canadian Council on Animal Care guidelines for humane animal use.
Informed consent was not obtained since human participants were not involved in the study.
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