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. 2018 May 11;17(5):1171–1180. doi: 10.1016/S2095-3119(17)61896-3

Identification of the strain-specifically truncated nonstructural protein 10 of porcine reproductive and respiratory syndrome virus in infected cells

Zhi-bang ZHANG 1,**,1, Lei XU 1,1, Xue-xia WEN 1, Jian-guo DONG 1, Lei ZHOU 1, Xin-na GE 1, Han-chun YANG 1,*, Xin GUO 1,***
PMCID: PMC7128467  PMID: 32288956

Abstract

The nonstructural protein 10 (nsp10) of porcine reproductive and respiratory syndrome virus (PRRSV) encodes for helicase which plays a vital role in viral replication. In the present study, a truncated form of nsp10, termed nsp10a, was found in PRRSV-infected cells and the production of nsp10a was strain-specific. Mass spectrometric analysis and deletion mutagenesis indicated that nsp10a may be short of about 70 amino acids in the N terminus of nsp10. Further studies by rescuing recombinant viruses showed that the Glu-69 in nsp10 was the key amino acid for nsp10a production. Finally, we demonstrated that nsp10a exerted little influence on the growth kinetics of PRRSV in vitro.

Keywords: porcine reproductive and respiratory syndrome virus (PRRSV), nonstructural protein 10 (nsp10), viral replication

Contributor Information

Zhi-bang ZHANG, Email: zhibangzhang@126.com.

Lei XU, Email: 358736128@qq.com.

Han-chun YANG, Email: yanghanchun1@cau.edu.cn.

Xin GUO, Email: guoxincau@cau.edu.cn.

References

  1. Bautista E M, Faaberg K S, Mickelson D, McGruder E D. Functional properties of the predicted helicase of porcine reproductive and respiratory syndrome virus. Virology. 2002;298:258–270. doi: 10.1006/viro.2002.1495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. den Boon J A, Snijder E J, Chirnside E D, de Vries A A, Horzinek M C, Spaan W J. Equine arteritis virus is not a togavirus but belongs to the coronaviruslike superfamily. Journal of Virology. 1991;65:2910–2920. doi: 10.1128/jvi.65.6.2910-2920.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Carter K L, Roizman B. The promoter and transcriptional unit of a novel herpes simplex virus 1 alpha gene are contained in, and encode a protein in frame with, the open reading frame of the alpha 22 gene. Journal of Virology. 1996;70:172–178. doi: 10.1128/jvi.70.1.172-178.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chen N, Cao Z, Yu X, Deng X, Zhao T, Wang L, Liu Q, Li X, Tian K. Emergence of novel European genotype porcine reproductive and respiratory syndrome virus in mainland China. Journal of General Virology. 2011;92:880–892. doi: 10.1099/vir.0.027995-0. [DOI] [PubMed] [Google Scholar]
  5. Deng Z, Lehmann K C, Li X, Feng C, Wang G, Zhang Q, Qi X, Yu L, Zhang X, Feng W, Wu W, Gong P, Tao Y, Posthuma C C, Snijder E J, Gorbalenya A E, Chen Z. Structural basis for the regulatory function of a complex zinc-binding domain in a replicative arterivirus helicase resembling a nonsense-mediated mRNA decay helicase. Nucleic Acids Research. 2014;42:3464–3477. doi: 10.1093/nar/gkt1310. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. van Dinten L C, van Tol H, Gorbalenya A E, Snijder E J. The predicted metal-binding region of the arterivirus helicase protein is involved in subgenomic mRNA synthesis, genome replication, and virion biogenesis. Journal of Virology. 2000;74:5213–5223. doi: 10.1128/jvi.74.11.5213-5223.2000. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Du J, Ge X, Dong H, Zhang N, Zhou L, Guo X, Yang H. The cellular interactome for glycoprotein 5 of the Chinese highly pathogenic porcine reproductive and respiratory syndrome virus. Journal of Integrative Agriculture. 2016;15:1833–1845. [Google Scholar]
  8. Fang P, Fang L, Hong Y, Liu X, Dong N, Ma P, Bi J, Wang D, Xiao S. Discovery of a novel accessory protein NS7a encoded by porcine deltacoronavirus. Journal of General Virology. 2017;98:173–178. doi: 10.1099/jgv.0.000690. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fang Y, Treffers E E, Li Y, Tas A, Sun Z, van der Meer Y, de Ru A H, van Veelen P A, Atkins J F, Snijder E J, Firth A E. Efficient –2 frameshifting by mammalian ribosomes to synthesize an additional arterivirus protein. Proceedings of the National Academy of Sciences of the United States of America. 2012;109:2920–2928. doi: 10.1073/pnas.1211145109. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Firth A E, Zevenhoven-Dobbe J C, Wills N M, Go Y Y, Balasuriya U B, Atkins J F, Snijder E J, Posthuma C C. Discovery of a small arterivirus gene that overlaps the GP5 coding sequence and is important for virus production. Journal of General Virology. 2011;92:1097–1106. doi: 10.1099/vir.0.029264-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Han J, Rutherford M S, Faaberg K S. Proteolytic products of the porcine reproductive and respiratory syndrome virus nsp2 replicase protein. Journal of Virology. 2010;84:10102–10112. doi: 10.1128/JVI.01208-10. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Holtkamp D J, Kliebenstein J B, Neumann E J, Zimmerman J J, Rotto H F, Yoder T K, Wang C, Yeske P, Mowrer C L, Haley C. Assessment of the economic impact of porcine reproductive and respiratory syndrome virus on United States pork producers. Journal of Swine Health and Production. 2013;21:72–84. [Google Scholar]
  13. Johnson C R, Griggs T F, Gnanandarajah J, Murtaugh M P. Novel structural protein in porcine reproductive and respiratory syndrome virus encoded by an alternative ORF5 present in all arteriviruses. Journal of General Virology. 2011;92:1107–1116. doi: 10.1099/vir.0.030213-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Li B, Fang L, Guo X, Gao J, Song T, Bi J, He K, Chen H, Xiao S. Epidemiology and evolutionary characteristics of the porcine reproductive and respiratory syndrome virus in China between 2006 and 2010. Journal of Clinical Microbiology. 2011;49:3175–3183. doi: 10.1128/JCM.00234-11. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Li Y, Tas A, Snijder E J, Fang Y. Identification of porcine reproductive and respiratory syndrome virus ORF1a-encoded non-structural proteins in virus-infected cells. Journal of General Virology. 2012;93:829–839. doi: 10.1099/vir.0.039289-0. [DOI] [PubMed] [Google Scholar]
  16. Li Y, Zhou L, Zhang J, Ge X, Zhou R, Zheng H, Geng G, Guo X, Yang H. Nsp9 and nsp10 contribute to the fatal virulence of highly pathogenic porcine reproductive and respiratory syndrome virus emerging in China. PLoS Pathogens. 2014;10:1004216. doi: 10.1371/journal.ppat.1004216. 1004216. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Liu F Y, Roizman B. The herpes simplex virus 1 gene encoding a protease also contains within its coding domain the gene encoding the more abundant substrate. Journal of virology. 1991;65:5149–5156. doi: 10.1128/jvi.65.10.5149-5156.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Liu F Y, Roizman B. The promoter, transcriptional unit, and coding sequence of herpes simplex virus 1 family 35 proteins are contained within and in frame with the UL26 open reading frame. Journal of Virology. 1991;65:206–212. doi: 10.1128/jvi.65.1.206-212.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Meng X J, Paul P S, Halbur P G. Molecular cloning and nucleotide sequencing of the 3'-terminal genomic RNA of the porcine reproductive and respiratory syndrome virus. Journal of General Virology. 1994;75:1795–1801. doi: 10.1099/0022-1317-75-7-1795. [DOI] [PubMed] [Google Scholar]
  20. Meulenberg J J. PRRSV, the virus. Veterinary Research. 2000;31:11–21. doi: 10.1051/vetres:2000103. [DOI] [PubMed] [Google Scholar]
  21. Nelsen C J, Murtaugh M P, Faaberg K S. Porcine reproductive and respiratory syndrome virus comparison: Divergent evolution on two continents. Journal of Virology. 1999;73:270–280. doi: 10.1128/jvi.73.1.270-280.1999. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Pejsak Z, Stadejek T, Markowskadaniel I. Clinical signs and economic losses caused by porcine reproductive and respiratory syndrome virus in a large breeding farm. Veterinary Microbiology. 1997;55:317–322. doi: 10.1016/s0378-1135(96)01326-0. [DOI] [PubMed] [Google Scholar]
  23. Rascón-Castelo E, Burgara-Estrella A, Mateu E, Hernández J. Immunological features of the non-structural proteins of porcine reproductive and respiratory syndrome virus. Viruses. 2015;7:873–886. doi: 10.3390/v7030873. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Seybert A, van Dinten L C, Snijder E J, Ziebuhr J. Biochemical characterization of the equine arteritis virus helicase suggests a close functional relationship between arterivirus and coronavirus helicases. Journal of Virology. 2000;74:9586–9593. doi: 10.1128/jvi.74.20.9586-9593.2000. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Seybert A, Posthuma C C, van Dinten L C, Snijder E J, Gorbalenya A E, Ziebuhr J. A complex zinc finger controls the enzymatic activities of nidovirus helicases. Journal of Virology. 2005;79:696–704. doi: 10.1128/JVI.79.2.696-704.2005. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Shi Y, Li Y, Lei Y, Ye G, Shen Z, Sun L, Luo R, Wang D, Fu Z F, Xiao S, Peng G. A dimerization-dependent mechanism drives the endoribonuclease function of porcine reproductive and respiratory syndrome virus nsp11. Journal of Virology. 2016;90:4579–4592. doi: 10.1128/JVI.03065-15. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Snijder E J, Kikkert M, Fang Y. Arterivirus molecular biology and pathogenesis. Journal of General Virology. 2013;94:2141–2163. doi: 10.1099/vir.0.056341-0. [DOI] [PubMed] [Google Scholar]
  28. Snijder E J, Meulenberg J J. The molecular biology of arteriviruses. Journal of General Virology. 1998;79:961–979. doi: 10.1099/0022-1317-79-5-961. [DOI] [PubMed] [Google Scholar]
  29. Tian K, Yu X, Zhao T, Feng Y, Cao Z, Wang C, Hu Y, Chen X, Hu D, Tian X, Liu D, Zhang S, Deng X, Ding Y, Yang L, Zhang Y, Xiao H, Qiao M, Wang B. Emergence of fatal PRRSV variants: Unparalleled outbreaks of atypical PRRS in China and molecular dissection of the unique hallmark. PLoS ONE. 2007;2:e526. doi: 10.1371/journal.pone.0000526. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Tong G, Zhou Y, Hao X, Tian Z, An T, Qiu H. Highly pathogenic porcine reproductive and respiratory syndrome, China. Emerging Infectious Diseases. 2007;13:1434–1436. doi: 10.3201/eid1309.070399. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Xie J, Cui T, Cui J, Chen Y, Zhang M, Zhou P, Deng S, Su S, Zhang G. Epidemiological and evolutionary characteristics of the PRRSV in Southern China from 2010 to 2013. Microbial Pathogenesis. 2014;75:7–15. doi: 10.1016/j.micpath.2014.08.001. [DOI] [PubMed] [Google Scholar]
  32. Zhang H, Guo X, Ge X, Chen Y, Sun Q, Yang H. Changes in the cellular proteins of pulmonary alveolar macrophage infected with porcine reproductive and respiratory syndrome virus by proteomics analysis. Journal of Proteome Research. 2009;8:3091–3097. doi: 10.1021/pr900002f. [DOI] [PubMed] [Google Scholar]
  33. Zhang Y, Li H, Peng G, Zhang Y, Gao X, Xiao S, Cao S, Chen H, Song Y. Mutational analysis of the functional sites in porcine reproductive and respiratory syndrome virus non-structural protein 10. Journal of General Virology. 2015;96:547–552. doi: 10.1099/jgv.0.000004. [DOI] [PubMed] [Google Scholar]
  34. Zhang Z, Wen X, Dong J, Ge X, Zhou L, Yang H, Guo X. Epitope mapping and characterization of a novel nsp10-specific monoclonal antibody that differentiates genotype 2 PRRSV from genotype 1 PRRSV. Virology Journal. 2017;14:116. doi: 10.1186/s12985-017-0782-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Zhao K, Ye C, Chang X, Jiang C, Wang S, Cai X, Tong G, Tian Z, Shi M, An T. Importation and recombination are responsible for the latest emergence of highly pathogenic PRRSV in China. Journal of virology. 2015;89:10712–10716. doi: 10.1128/JVI.01446-15. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Zhou L, Wang Z, Ding Y, Ge X, Guo X, Yang H. NADC30-like strain of porcine reproductive and respiratory syndrome virus, China. Emerging Infectious Diseases. 2015;21:2256–2257. doi: 10.3201/eid2112.150360. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Zhou L, Yang H. Porcine reproductive and respiratory syndrome in China. Virus Research. 2010;154:31–37. doi: 10.1016/j.virusres.2010.07.016. [DOI] [PubMed] [Google Scholar]
  38. Zhou L, Zhang J, Zeng J, Yin S, Li Y, Zheng L, Guo X, Ge X, Yang H. The 30-amino-acid deletion in the Nsp2 of highly pathogenic porcine reproductive and respiratory syndrome virus emerging in China is not related to its virulence. Journal of Virology. 2009;83:5156–5167. doi: 10.1128/JVI.02678-08. [DOI] [PMC free article] [PubMed] [Google Scholar]

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