uS10, a novel N-interacting protein, inhibits classical swine fever virus replication Free

Abstract

Classical swine fever (CSF) is a severe, febrile and highly contagious disease caused by classical swine fever virus (CSFV) that has resulted in huge economic losses in the pig industry worldwide. CSFV N has been actively studied but remains incompletely understood. Few studies have investigated the cellular proteins that interact with N and their participation in viral replication. Here, the yeast two-hybrid (Y2H) system was employed to screen N-interacting proteins from a porcine alveolar macrophage (PAM) cDNA library, and a search of the NCBI database revealed that 15 cellular proteins interact with N. The interaction of N with ribosomal protein S20, also known as universal S10 (uS10), was further confirmed by co-immunoprecipitation and glutathione -transferase pull-down assays. Furthermore, uS10 overexpression inhibited CSFV replication, whereas the knockdown of uS10 promoted CSFV replication in PAMs. In addition, N or CSFV reduced uS10 expression in PAMs in a proteasome-dependent manner, indicating that N–uS10 interaction might contribute to persistent CSFV replication. Our previous research showed that CSFV decreases Toll-like receptor 3 (TLR3) expression. The results showed that uS10 knockdown reduced TLR3 expression, and that uS10 overexpression increased TLR3 expression. Notably, uS10 knockdown did not promote CSFV replication following TLR3 overexpression. Conversely, uS10 overexpression did not inhibit CSFV replication following TLR3 knockdown. These results revealed that uS10 inhibits CSFV replication by modulating TLR3 expression. This work addresses a novel aspect of the regulation of the innate antiviral immune response during CSFV infection.

Keyword(s): CSFV , Npro , replication , TLR3 and uS10
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2017-07-01
2024-03-29
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References

  1. Simmonds P, Becher P, Bukh J, Gould EA, Meyers G et al. ICTV virus taxonomy profile: Flaviviridae. J Gen Virol 2017; 98:2–3 [View Article][PubMed]
    [Google Scholar]
  2. Lamp B, Riedel C, Wentz E, Tortorici MA, Rümenapf T. Autocatalytic cleavage within classical swine fever virus NS3 leads to a functional separation of protease and helicase. J Virol 2013; 87:11872–11883 [View Article][PubMed]
    [Google Scholar]
  3. Lindenbach BD, Rice CM. Flavivirus: the viruses and their replication. In Knipe DM, Howley PM. (editors) Fields Virology, 5th ed. Philadelphia: Lippincott Williams & WilkinsPA; 2007 pp. 1101–1152
    [Google Scholar]
  4. Rümenapf T, Thiel HJ. Molecular biology of pestiviruses. In Mettenleiter T, Sobrino F. (editors) Animal Viruses: Molecular Biology UK: Caister Academic Press; 2008 pp. 39–96
    [Google Scholar]
  5. La Rocca SA, Herbert RJ, Crooke H, Drew TW, Wileman TE et al. Loss of interferon regulatory factor 3 in cells infected with classical swine fever virus involves the N-terminal protease, Npro. J Virol 2005; 79:7239–7247 [View Article][PubMed]
    [Google Scholar]
  6. Bauhofer O, Summerfield A, Sakoda Y, Tratschin JD, Hofmann MA et al. Classical swine fever virus Nprointeracts with interferon regulatory factor 3 and induces its proteasomal degradation. J Virol 2007; 81:3087–3096 [View Article][PubMed]
    [Google Scholar]
  7. Mine J, Tamura T, Mitsuhashi K, Okamatsu M, Parchariyanon S et al. The N-terminal domain of Npro of classical swine fever virus determines its stability and regulates type I IFN production. J Gen Virol 2015; 96:1746–1756 [View Article][PubMed]
    [Google Scholar]
  8. Seago J, Hilton L, Reid E, Doceul V, Jeyatheesan J et al. The Npro product of classical swine fever virus and bovine viral diarrhea virus uses a conserved mechanism to target interferon regulatory factor-3. J Gen Virol 2007; 88:3002–3006 [View Article][PubMed]
    [Google Scholar]
  9. Seago J, Goodbourn S, Charleston B. The classical swine fever virus Npro product is degraded by cellular proteasomes in a manner that does not require interaction with interferon regulatory factor 3. J Gen Virol 2010; 91:721–726 [View Article][PubMed]
    [Google Scholar]
  10. Ruggli N, Summerfield A, Fiebach AR, Guzylack-Piriou L, Bauhofer O et al. Classical swine fever virus can remain virulent after specific elimination of the interferon regulatory factor 3-degrading function of Npro. J Virol 2009; 83:817–829 [View Article][PubMed]
    [Google Scholar]
  11. Tamura T, Nagashima N, Ruggli N, Summerfield A, Kida H et al. Npro of classical swine fever virus contributes to pathogenicity in pigs by preventing type I interferon induction at local replication sites. Vet Res 2014; 45:47 [View Article][PubMed]
    [Google Scholar]
  12. Fiebach AR, Guzylack-Piriou L, Python S, Summerfield A, Ruggli N. Classical swine fever virus Npro limits type I interferon induction in plasmacytoid dendritic cells by interacting with interferon regulatory factor 7. J Virol 2011; 85:8002–8011 [View Article][PubMed]
    [Google Scholar]
  13. Doceul V, Charleston B, Crooke H, Reid E, Powell PP et al. The Npro product of classical swine fever virus interacts with IκBα, the NF-κB inhibitor. J Gen Virol 2008; 89:1881–1889 [View Article][PubMed]
    [Google Scholar]
  14. Chen FW, Ioannou YA. Ribosomal proteins in cell proliferation and apoptosis. Int Rev Immunol 1999; 18:429–448 [View Article][PubMed]
    [Google Scholar]
  15. Wang G, Inaoka T, Okamoto S, Ochi K. A novel insertion mutation in Streptomyces coelicolor ribosomal S12 protein results in paromomycin resistance and antibiotic overproduction. Antimicrob Agents Chemother 2009; 53:1019–1026 [View Article][PubMed]
    [Google Scholar]
  16. Majzoub K, Hafirassou ML, Meignin C, Goto A, Marzi S et al. RACK1 controls IRES-mediated translation of viruses. Cell 2014; 159:1086–1095 [View Article][PubMed]
    [Google Scholar]
  17. Fuchs G, Petrov AN, Marceau CD, Popov LM, Chen J et al. Kinetic pathway of 40S ribosomal subunit recruitment to hepatitis C virus internal ribosome entry site. Proc Natl Acad Sci USA 2015; 112:319–325 [View Article][PubMed]
    [Google Scholar]
  18. Carvajal F, Vallejos M, Walters B, Contreras N, Hertz MI et al. Structural domains within the HIV-1 mRNA and the ribosomal protein S25 influence cap-independent translation initiation. FEBS J 2016; 283:2508–2527 [View Article][PubMed]
    [Google Scholar]
  19. Jefferson M, Donaszi-Ivanov A, Pollen S, Dalmay T, Saalbach G et al. Host factors that interact with the pestivirus N-terminal protease, Npro, are components of the ribonucleoprotein complex. J Virol 2014; 88:10340–10353 [View Article][PubMed]
    [Google Scholar]
  20. de Bortoli M, Castellino RC, Lu XY, Deyo J, Sturla LM et al. Medulloblastoma outcome is adversely associated with overexpression of EEF1D, RPL30, and RPS20 on the long arm of chromosome 8. BMC Cancer 2006; 6:223 [View Article][PubMed]
    [Google Scholar]
  21. Yan X, Xie J, Li J, Shuanghu C, Wu Z et al. Screening and analysis on the protein interaction of the protein VP7 in grass carp reovirus. Virus Genes 2015; 50:425–433 [View Article][PubMed]
    [Google Scholar]
  22. Cao Z, Guo K, Zheng M, Ning P, Li H et al. A comparison of the impact of Shimen and C strains of classical swine fever virus on Toll-like receptor expression. J Gen Virol 2015; 96:1732–1745 [View Article][PubMed]
    [Google Scholar]
  23. Brückner A, Polge C, Lentze N, Auerbach D, Schlattner U. Yeast two-hybrid, a powerful tool for systems biology. Int J Mol Sci 2009; 10:2763–2788 [View Article][PubMed]
    [Google Scholar]
  24. Mcgowan KA, Li JZ, Park CY, Beaudry V, Tabor HK et al. Ribosomal mutations cause p53-mediated dark skin and pleiotropic effects. Nat Genet 2008; 40:963–970 [View Article][PubMed]
    [Google Scholar]
  25. Kemp EH, Herd LM, Waterman EA, Wilson AG, Weetman AP et al. Immunoscreening of phage-displayed cDNA-encoded polypeptides identifies B cell targets in autoimmune disease. Biochem Biophys Res Commun 2002; 298:169–177 [View Article][PubMed]
    [Google Scholar]
  26. Nieminen TT, O'Donohue MF, Wu Y, Lohi H, Scherer SW et al. Germline mutation of RPS20, encoding a ribosomal protein, causes predisposition to hereditary nonpolyposis colorectal carcinoma without DNA mismatch repair deficiency. Gastroenterology 2014; 147:595–598 [View Article][PubMed]
    [Google Scholar]
  27. Goldstone SD, Lavin MF. Isolation of a cDNA clone, encoding the ribosomal protein S20, downregulated during the onset of apoptosis in a human leukaemic cell line. Biochem Biophys Res Commun 1993; 196:619–623 [View Article][PubMed]
    [Google Scholar]
  28. Yu H, Yao LH, Chen AJ, He J, Jia RQ et al. [Screening for new binding proteins which interact with BM2 of influenza B virus with yeast two-hybrid system]. Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi 2005; 19:182–184[PubMed]
    [Google Scholar]
  29. Daftuar L, Zhu Y, Jacq X, Prives C. Ribosomal proteins RPL37, RPS15 and RPS20 regulate the Mdm2-p53-MdmX network. PLoS One 2013; 8:e68667 [View Article][PubMed]
    [Google Scholar]
  30. Bauhofer O, Summerfield A, Sakoda Y, Tratschin JD, Hofmann MA et al. Classical swine fever virus Npro interacts with interferon regulatory factor 3 and induces its proteasomal degradation. J Virol 2007; 81:3087–3096 [View Article][PubMed]
    [Google Scholar]
  31. Fiebach AR, Guzylack-Piriou L, Python S, Summerfield A, Ruggli N. Classical swine fever virus Npro limits type I interferon induction in plasmacytoid dendritic cells by interacting with interferon regulatory factor 7. J Virol 2011; 85:8002–8011 [View Article][PubMed]
    [Google Scholar]
  32. Zhang C, Kang K, Ning P, Peng Y, Lin Z et al. Heat shock protein 70 is associated with CSFV NS5A protein and enhances viral RNA replication. Virology 2015; 482:9–18 [View Article][PubMed]
    [Google Scholar]
  33. Yang Z, Shi Z, Guo H, Qu H, Zhang Y et al. Annexin 2 is a host protein binding to classical swine fever virus E2 glycoprotein and promoting viral growth in PK-15 cells. Virus Res 2015; 201:16–23 [View Article][PubMed]
    [Google Scholar]
  34. Li S, Wang J, He WR, Feng S, Li Y et al. Thioredoxin 2 is a novel E2-interacting protein that inhibits the replication of classical swine fever virus. J Virol 2015; 89:8510–8524 [View Article][PubMed]
    [Google Scholar]
  35. Wang J, Chen S, Liao Y, Zhang E, Feng S et al. Mitogen-activated protein kinase kinase 2 (MEK2), a novel E2-interacting protein, promotes the growth of classical swine fever virus via attenuation of the JAK-STAT signaling pathway. J Virol 2016; 90:10271–10283 [View Article]
    [Google Scholar]
  36. Abe T, Kaname Y, Hamamoto I, Tsuda Y, Wen X et al. Hepatitis C virus nonstructural protein 5A modulates the toll-like receptor-MyD88-dependent signaling pathway in macrophage cell lines. J Virol 2007; 81:8953–8966 [View Article][PubMed]
    [Google Scholar]
  37. Akira S, Uematsu S, Takeuchi O. Pathogen recognition and innate immunity. Cell 2006; 124:783–801 [View Article][PubMed]
    [Google Scholar]
  38. Schulz O, Diebold SS, Chen M, Näslund TI, Nolte MA et al. Toll-like receptor 3 promotes cross-priming to virus-infected cells. Nature 2005; 433:887–892 [View Article][PubMed]
    [Google Scholar]
  39. Nasirudeen AM, Wong HH, Thien P, Xu S, Lam KP et al. RIG-I, MDA5 and TLR3 synergistically play an important role in restriction of dengue virus infection. PLoS Negl Trop Dis 2011; 5:e926 [View Article][PubMed]
    [Google Scholar]
  40. Wong JP, Christopher ME, Viswanathan S, Dai X, Salazar AM et al. Antiviral role of toll-like receptor-3 agonists against seasonal and avian influenza viruses. Curr Pharm Des 2009; 15:1269–1274 [View Article][PubMed]
    [Google Scholar]
  41. Edelmann KH, Richardson-Burns S, Alexopoulou L, Tyler KL, Flavell RA et al. Does Toll-like receptor 3 play a biological role in virus infections?. Virology 2004; 322:231–238 [View Article][PubMed]
    [Google Scholar]
  42. Wang T, Town T, Alexopoulou L, Anderson JF, Fikrig E et al. Toll-like receptor 3 mediates West Nile virus entry into the brain causing lethal encephalitis. Nat Med 2004; 10:1366–1373 [View Article][PubMed]
    [Google Scholar]
  43. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT Method. Methods 2001; 25:402–408 [View Article][PubMed]
    [Google Scholar]
  44. Szklarczyk D, Franceschini A, Kuhn M, Simonovic M, Roth A et al. The STRING database in 2011: functional interaction networks of proteins, globally integrated and scored. Nucleic Acids Res 2011; 39:D561–D568 [View Article][PubMed]
    [Google Scholar]
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