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Characterization of the interactome of the porcine reproductive and respiratory syndrome virus glycoprotein-5

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Abstract

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.

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References

  1. Adams MJ, Lefkowitz EJ, King AM, Harrach B, Harrison RL, Knowles NJ, Kropinski AM, Krupovic M, Kuhn JH, Mushegian AR, Nibert M, Sabanadzovic S, Sanfacon H, Siddell SG, Simmonds P, Varsani A, Zerbini FM, Gorbalenya AE, Davison AJ (2016) Ratification vote on taxonomic proposals to the International Committee on Taxonomy of Viruses (2016). Arch Virol 161:2921–2949

    Article  PubMed  CAS  Google Scholar 

  2. Cervantes-Salazar M, Angel-Ambrocio AH, Soto-Acosta R, Bautista-Carbajal P, Hurtado-Monzon AM, Alcaraz-Estrada SL, Ludert JE, Del Angel RM (2015) Dengue virus NS1 protein interacts with the ribosomal protein RPL18: this interaction is required for viral translation and replication in Huh-7 cells. Virology 484:113–126

    Article  PubMed  CAS  Google Scholar 

  3. Chartier C, Degryse E, Gantzer M, Dieterle A, Pavirani A, Mehtali M (1996) Efficient generation of recombinant adenovirus vectors by homologous recombination in Escherichia coli. J Virol 70:4805–4810

    PubMed  PubMed Central  CAS  Google Scholar 

  4. Chen Y, Lu Z, Zhang L, Gao L, Wang N, Gao X, Wang Y, Li K, Gao Y, Cui H, Gao H, Liu C, Zhang Y, Qi X, Wang X (2016) Ribosomal protein L4 interacts with viral protein VP3 and regulates the replication of infectious bursal disease virus. Virus Res 211:73–78

    Article  PubMed  CAS  Google Scholar 

  5. Delputte PL, Nauwynck HJ (2004) Porcine arterivirus infection of alveolar macrophages is mediated by sialic acid on the virus. J Virol 78:8094–8101

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  6. Dong S, Liu L, Wu W, Armstrong SD, Xia D, Nan H, Hiscox JA, Chen H (2016) Determination of the interactome of non-structural protein12 from highly pathogenic porcine reproductive and respiratory syndrome virus with host cellular proteins using high throughput proteomics and identification of HSP70 as a cellular factor for virus replication. J Proteom 146:58–69

    Article  CAS  Google Scholar 

  7. Fang Y, Treffers EE, Li Y, Tas A, Sun Z, van der Meer Y, de Ru AH, van Veelen PA, Atkins JF, Snijder EJ, Firth AE (2012) Efficient -2 frameshifting by mammalian ribosomes to synthesize an additional arterivirus protein. Proc Natl Acad Sci USA 109:E2920–2928

    Article  PubMed  PubMed Central  Google Scholar 

  8. Gao J, Ji P, Zhang M, Wang X, Li N, Wang C, Xiao S, Mu Y, Zhao Q, Du T, Sun Y, Hiscox JA, Zhang G, Zhou EM (2014) GP5 expression in Marc-145 cells inhibits porcine reproductive and respiratory syndrome virus infection by inducing beta interferon activity. Vet Microbiol 174:409–418

    Article  PubMed  CAS  Google Scholar 

  9. Gao J, Xiao S, Xiao Y, Wang X, Zhang C, Zhao Q, Nan Y, Huang B, Liu H, Liu N, Lv J, Du T, Sun Y, Mu Y, Wang G, Syed SF, Zhang G, Hiscox JA, Goodfellow I, Zhou EM (2016) MYH9 is an essential factor for porcine reproductive and respiratory syndrome virus infection. Sci Rep 6:25120

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  10. Johnson CR, Griggs TF, Gnanandarajah J, Murtaugh MP (2011) Novel structural protein in porcine reproductive and respiratory syndrome virus encoded by an alternative ORF5 present in all arteriviruses. J Gen Virol 92:1107–1116

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  11. Jourdan SS, Osorio F, Hiscox JA (2012) An interactome map of the nucleocapsid protein from a highly pathogenic North American porcine reproductive and respiratory syndrome virus strain generated using SILAC-based quantitative proteomics. Proteomics 12:1015–1023

    Article  PubMed  CAS  Google Scholar 

  12. Lee C, Bachand A, Murtaugh MP, Yoo D (2004) Differential host cell gene expression regulated by the porcine reproductive and respiratory syndrome virus GP4 and GP5 glycoproteins. Vet Immunol Immunopathol 102:189–198

    Article  PubMed  CAS  Google Scholar 

  13. Lee C, Rogan D, Erickson L, Zhang J, Yoo D (2004) Characterization of the porcine reproductive and respiratory syndrome virus glycoprotein 5 (GP5) in stably expressing cells. Virus Res 104:33–38

    Article  PubMed  CAS  Google Scholar 

  14. Liu L, Lear Z, Hughes DJ, Wu W, Zhou EM, Whitehouse A, Chen H, Hiscox JA (2015) 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. Vet Microbiol 176:109–119

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  15. Lunney JK, Fang Y, Ladinig A, Chen N, Li Y, Rowland B, Renukaradhya GJ (2016) Porcine reproductive and respiratory syndrome virus (PRRSV): pathogenesis and Interaction with the Immune System. Annu Rev Anim Biosci 4:129–154

    Article  PubMed  CAS  Google Scholar 

  16. Mardassi H, Massie B, Dea S (1996) Intracellular synthesis, processing, and transport of proteins encoded by ORFs 5 to 7 of porcine reproductive and respiratory syndrome virus. Virology 221:98–112

    Article  PubMed  CAS  Google Scholar 

  17. Mu Y, Li L, Zhang B, Huang B, Gao J, Wang X, Wang C, Xiao S, Zhao Q, Sun Y, Zhang G, Hiscox JA, Zhou EM (2015) Glycoprotein 5 of porcine reproductive and respiratory syndrome virus strain SD16 inhibits viral replication and causes G2/M cell cycle arrest, but does not induce cellular apoptosis in Marc-145 cells. Virology 484:136–145

    Article  PubMed  CAS  Google Scholar 

  18. Nelsen CJ, Murtaugh MP, Faaberg KS (1999) Porcine reproductive and respiratory syndrome virus comparison: divergent evolution on two continents. J Virol 73:270–280

    PubMed  PubMed Central  CAS  Google Scholar 

  19. Nelson EA, Christopher-Hennings J, Drew T, Wensvoort G, Collins JE, Benfield DA (1993) Differentiation of U.S. and European isolates of porcine reproductive and respiratory syndrome virus by monoclonal antibodies. J Clin Microbiol 31:3184–3189

    PubMed  PubMed Central  CAS  Google Scholar 

  20. Ni YY, Huang YW, Cao D, Opriessnig T, Meng XJ (2011) Establishment of a DNA-launched infectious clone for a highly pneumovirulent strain of type 2 porcine reproductive and respiratory syndrome virus: identification and in vitro and in vivo characterization of a large spontaneous deletion in the nsp2 region. Virus Res 160:264–273

    Article  PubMed  CAS  Google Scholar 

  21. Ostrowski M, Galeota JA, Jar AM, Platt KB, Osorio FA, Lopez OJ (2002) Identification of neutralizing and nonneutralizing epitopes in the porcine reproductive and respiratory syndrome virus GP5 ectodomain. J Virol 76:4241–4250

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  22. Pujhari S, Kryworuchko M, Zakhartchouk AN (2014) Role of phosphatidylinositol-3-kinase (PI3K) and the mammalian target of rapamycin (mTOR) signalling pathways in porcine reproductive and respiratory syndrome virus (PRRSV) replication. Virus Res 194:138–144

    Article  PubMed  CAS  Google Scholar 

  23. Pujhari S, Zakhartchouk AN (2016) Porcine reproductive and respiratory syndrome virus envelope (E) protein interacts with mitochondrial proteins and induces apoptosis. Arch Virol 161:1821–1830

    Article  PubMed  CAS  Google Scholar 

  24. Snijder EJ, Kikkert M, Fang Y (2013) Arterivirus molecular biology and pathogenesis. J Gen Virol 94:2141–2163

    Article  PubMed  CAS  Google Scholar 

  25. Song S, Bi J, Wang D, Fang L, Zhang L, Li F, Chen H, Xiao S (2013) Porcine reproductive and respiratory syndrome virus infection activates IL-10 production through NF-kappaB and p38 MAPK pathways in porcine alveolar macrophages. Dev Comp Immunol 39:265–272

    Article  PubMed  CAS  Google Scholar 

  26. Song T, Fang L, Wang D, Zhang R, Zeng S, An K, Chen H, Xiao S (2016) Quantitative interactome reveals that porcine reproductive and respiratory syndrome virus nonstructural protein 2 forms a complex with viral nucleocapsid protein and cellular vimentin. J Proteom 142:70–81

    Article  CAS  Google Scholar 

  27. Suarez P, Diaz-Guerra M, Prieto C, Esteban M, Castro JM, Nieto A, Ortin J (1996) Open reading frame 5 of porcine reproductive and respiratory syndrome virus as a cause of virus-induced apoptosis. J Virol 70:2876–2882

    PubMed  PubMed Central  CAS  Google Scholar 

  28. Sun L, Li Y, Liu R, Wang X, Gao F, Lin T, Huang T, Yao H, Tong G, Fan H, Wei Z, Yuan S (2013) Porcine reproductive and respiratory syndrome virus ORF5a protein is essential for virus viability. Virus Res 171:178–185

    Article  PubMed  CAS  Google Scholar 

  29. Thaa B, Sinhadri BC, Tielesch C, Krause E, Veit M (2013) Signal peptide cleavage from GP5 of PRRSV: a minor fraction of molecules retains the decoy epitope, a presumed molecular cause for viral persistence. PloS One 8:e65548

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  30. van Marle G, Dobbe JC, Gultyaev AP, Luytjes W, Spaan WJ, Snijder EJ (1999) Arterivirus discontinuous mRNA transcription is guided by base pairing between sense and antisense transcription-regulating sequences. Proc Natl Acad Sci USA 96:12056–12061

    Article  PubMed  PubMed Central  Google Scholar 

  31. Wang L, Zhou L, Zhang H, Li Y, Ge X, Guo X, Yu K, Yang H (2014) Interactome profile of the host cellular proteins and the nonstructural protein 2 of porcine reproductive and respiratory syndrome virus. PloS One 9:e99176

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  32. Wang Q, Li Y, Dong H, Wang L, Peng J, An T, Yang X, Tian Z, Cai X (2017) Identification of host cellular proteins that interact with the M protein of a highly pathogenic porcine reproductive and respiratory syndrome virus vaccine strain. Virol J 14:39

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  33. Wissink EH, Kroese MV, van Wijk HA, Rijsewijk FA, Meulenberg JJ, Rottier PJ (2005) Envelope protein requirements for the assembly of infectious virions of porcine reproductive and respiratory syndrome virus. J Virol 79:12495–12506

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  34. Wu WH, Fang Y, Farwell R, Steffen-Bien M, Rowland RR, Christopher-Hennings J, Nelson EA (2001) A 10-kDa structural protein of porcine reproductive and respiratory syndrome virus encoded by ORF2b. Virology 287:183–191

    Article  PubMed  CAS  Google Scholar 

  35. Xiao Y, Wu W, Gao J, Smith N, Burkard C, Xia D, Zhang M, Wang C, Archibald A, Digard P, Zhou EM, Hiscox JA (2016) 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. J Proteome Res 15:1388–1401

    Article  PubMed  CAS  Google Scholar 

  36. Yang L, Zhang YJ (2016) Antagonizing cytokine-mediated JAK-STAT signaling by porcine reproductive and respiratory syndrome virus. Vet Microbiol 209:57–65

    Article  PubMed  CAS  Google Scholar 

  37. Zhu L, Yang S, Tong W, Zhu J, Yu H, Zhou Y, Morrison RB, Tong G (2013) Control of the PI3K/Akt pathway by porcine reproductive and respiratory syndrome virus. Arch Virol 158:1227–1234

    Article  PubMed  CAS  Google Scholar 

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Acknowledgements

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.

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Author notes

  1. Maodong Zhang

    Present address: Department of Veterinary Pathology, University of Saskatchewan, 52 Campus Drive, Saskatoon, SK, S7N 5B4, Canada

  2. Alexander Zakhartchouk

    Present address: Department of Veterinary Microbiology, Western College of Veterinary Medicine, University of Saskatchewan, 52 Campus Drive, Saskatoon, SK, S7N 5B4, Canada

Authors and Affiliations

  1. Vaccine and Infectious Disease Organization-International Vaccine Center (VIDO-InterVac), University of Saskatchewan, 120 Veterinary Road, Saskatoon, SK, S7N 5E3, Canada

    Maodong Zhang & Alexander Zakhartchouk

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  1. Maodong Zhang
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Correspondence to Alexander Zakhartchouk.

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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.

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Informed consent was not obtained since human participants were not involved in the study.

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Handling Editor: Sheela Ramamoorthy.

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Zhang, M., Zakhartchouk, A. Characterization of the interactome of the porcine reproductive and respiratory syndrome virus glycoprotein-5. Arch Virol 163, 1595–1605 (2018). https://doi.org/10.1007/s00705-018-3787-9

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  • DOI: https://doi.org/10.1007/s00705-018-3787-9

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