The PDZ-Binding Motif of Severe Acute Respiratory Syndrome Coronavirus Envelope Protein Is a Determinant of Viral Pathogenesis

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SARS-CoV caused a worldwide epidemic infecting 8000 people with a mortality of about 10%. A recombinant SARS-CoV lacking the E protein was attenuated in vivo. The E protein contains a PDZ-binding motif (PBM), a domain potentially involved in the interaction with more than 400 cellular proteins, which highlights its relevance in modulating host-cell behavior. To analyze the contributions of this motif to virulence, recombinant viruses with or without E protein PBM were generated. Recombinant SARS-CoVs lacking E protein PBM caused minimal lung damage and were attenuated, in contrast to viruses containing this motif, indicating that E protein PBM is a virulence determinant. E protein PBM induces the deleterious exacerbated immune response triggered during SARS-CoV infection, and interacts with the cellular protein syntenin, as demonstrated using proteomic analyses. Interestingly, syntenin redistributed from nucleus to cytoplasm during SARS-CoV infection, activating p38 MAPK and triggering the overexpression of inflammatory cytokines. Furthermore, silencing of syntenin using siRNAs led to a decrease in p38 MAPK activation. In addition, administration of a p38 MAPK inhibitor led to an increase in mice survival after SARS-CoV infection. These results indicate that syntenin and p38 MAPK are potential therapeutic targets to reduce the exacerbated immune response during SARS-CoV infection.

Vyšlo v časopise: The PDZ-Binding Motif of Severe Acute Respiratory Syndrome Coronavirus Envelope Protein Is a Determinant of Viral Pathogenesis. PLoS Pathog 10(8): e32767. doi:10.1371/journal.ppat.1004320
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1004320

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1. PeirisJS, YuenKY, OsterhausAD, StohrK (2003) The severe acute respiratory syndrome. N Engl J Med 349: 2431–2441.

2. LauSK, WooPC, LiKS, HuangY, TsoiHW, et al. (2005) Severe acute respiratory syndrome coronavirus-like virus in Chinese horseshoe bats. Proc Natl Acad Sci USA 102: 14040–14045.

3. LiW, ShiZ, YuM, RenW, SmithC, et al. (2005) Bats are natural reservoirs of SARS-like coronaviruses. Science 310: 676–679.

4. WooPC, LauSK, LiKS, PoonRW, WongBH, et al. (2006) Molecular diversity of coronaviruses in bats. Virology 351: 180–187.

5. DominguezSR, O'SheaTJ, OkoLM, HolmesKV (2007) Detection of group 1 coronaviruses in bats in North America. Emerg Infect Dis 13: 1295–1300.

6. ZakiAM, van BoheemenS, BestebroerTM, OsterhausAD, FouchierRA (2012) Isolation of a novel coronavirus from a man with pneumonia in Saudi Arabia. N Engl J Med 367: 1814–1820.

7. BerminghamA, ChandM, BrownC, AaronsE, TongC, et al. (2012) Severe respiratory illness caused by a novel coronavirus, in a patient transferred to the United Kingdom from the Middle East, September 2012. Euro Surveill 17 pii: 20290.

8. AssiriA, McGeerA, PerlTM, PriceCS, Al RabeeahAA, et al. (2013) Hospital outbreak of Middle East respiratory syndrome coronavirus. N Engl J Med 369: 407–16.

9. Enjuanes L, DeDiego ML, Alvarez E, Capiscol C, Baric R (2008) Vaccines for severe acute respiratory syndrome virus and other coronaviruses. In: Perlman S, Gallagher TM, Snijder EJ, editors. Nidoviruses. Washington: ASM Press. pp. 379–408.

10. TorresJ, ParthasarathyK, LinX, SaravananR, LiuDX (2006) Model of a putative pore: the pentameric alpha-helical bundle of SARS coronavirus E protein in lipid bilayers. Biophys J 91: 938–947.

11. Verdia-BaguenaC, Nieto-TorresJL, AlcarazA, DediegoML, TorresJ, et al. (2012) Coronavirus E protein forms ion channels with functionally and structurally-involved membrane lipids. Virology 432: 485–494.

12. Verdia-BaguenaC, Nieto-TorresJL, AlcarazA, DediegoML, EnjuanesL, et al. (2013) Analysis of SARS-CoV E protein ion channel activity by tuning the protein and lipid charge. Biochim Biophys Acta 1828: 2026–2031.

13. Nieto-TorresJL, DediegoML, Verdia-BaguenaC, Jimenez-GuardenoJM, Regla-NavaJA, et al. (2014) Severe acute respiratory syndrome coronavirus envelope protein ion channel activity promotes virus fitness and pathogenesis. PLoS Pathog 10: e1004077.

14. NalB, ChanC, KienF, SiuL, TseJ, et al. (2005) Differential maturation and subcellular localization of severe acute respiratory syndrome coronavirus surface proteins S, M and E. J Gen Virol 86: 1423–1434.

15. Nieto-TorresJL, DediegoML, AlvarezE, Jimenez-GuardenoJM, Regla-NavaJA, et al. (2011) Subcellular location and topology of severe acute respiratory syndrome coronavirus envelope protein. Virology 415: 69–82.

16. RuchTR, MachamerCE (2012) A single polar residue and distinct membrane topologies impact the function of the infectious bronchitis coronavirus E protein. PLoS Pathog 8: e1002674.

17. YeY, HogueBG (2007) Role of the coronavirus E viroporin protein transmembrane domain in virus assembly. J Virol 81: 3597–3607.

18. DeDiegoML, AlvarezE, AlmazanF, RejasMT, LamirandeE, et al. (2007) A severe acute respiratory syndrome coronavirus that lacks the E gene is attenuated in vitro and in vivo. J Virol 81: 1701–1713.

19. DeDiegoML, PeweL, AlvarezE, RejasMT, PerlmanS, et al. (2008) Pathogenicity of severe acute respiratory coronavirus deletion mutants in hACE-2 transgenic mice. Virology 376: 379–389.

20. NetlandJ, DeDiegoML, ZhaoJ, FettC, AlvarezE, et al. (2010) Immunization with an attenuated severe acute respiratory syndrome coronavirus deleted in E protein protects against lethal respiratory disease. Virology 399: 120–128.

21. FettC, DeDiegoML, Regla-NavaJA, EnjuanesL, PerlmanS (2013) Complete protection against severe acute respiratory syndrome coronavirus-mediated lethal respiratory disease in aged mice by immunization with a mouse-adapted virus lacking E protein. J Virol 87: 6551–6559.

22. DeDiegoML, Nieto-TorresJL, Jimenez-GuardenoJM, Regla-NavaJA, AlvarezE, et al. (2011) Severe acute respiratory syndrome coronavirus envelope protein regulates cell stress response and apoptosis. PLoS pathog 7: e1002315.

23. TeohKT, SiuYL, ChanWL, SchluterMA, LiuCJ, et al. (2010) The SARS coronavirus E protein interacts with PALS1 and alters tight junction formation and epithelial morphogenesis. Mol Biol Cell 21: 3838–3852.

24. HarrisBZ, LimWA (2001) Mechanism and role of PDZ domains in signaling complex assembly. J Cell Sci 114: 3219–3231.

25. HungAY, ShengM (2002) PDZ domains: structural modules for protein complex assembly. J Biol Chem 277: 5699–5702.

26. MunzM, HeinJ, BigginPC (2012) The role of flexibility and conformational selection in the binding promiscuity of PDZ domains. PLoS Comput Biol 8: e1002749.

27. GerekZN, KeskinO, OzkanSB (2009) Identification of specificity and promiscuity of PDZ domain interactions through their dynamic behavior. Proteins 77: 796–811.

28. JavierRT, RiceAP (2011) Emerging theme: cellular PDZ proteins as common targets of pathogenic viruses. J Virol 85: 11544–11556.

29. PontingCP (1997) Evidence for PDZ domains in bacteria, yeast, and plants. Protein Sci 6: 464–468.

30. SpallerMR (2006) Act globally, think locally: systems biology addresses the PDZ domain. ACS Chem Biol 1: 207–210.

31. RobertsA, DemingD, PaddockCD, ChengA, YountB, et al. (2007) A mouse-adapted SARS-coronavirus causes disease and mortality in BALB/c mice. PLoS Pathog 3: 23–37.

32. DeDiegoML, Nieto-TorresJL, Regla-NavaJA, Jimenez-GuardenoJM, Fernandez-DelgadoR, et al. (2014) Inhibition of NF-kappaB mediated inflammation in severe acute respiratory syndome coronavirus-infected mice increases survival. J Virol 88: 913–924.

33. Regla-NavaJA, Jimenez-GuardenoJM, Nieto-TorresJL, GallagherTM, EnjuanesL, et al. (2013) The replication of a mouse adapted SARS-CoV in a mouse cell line stably expressing the murine SARS-CoV receptor mACE2 efficiently induces the expression of proinflammatory cytokines. J Virol Methods 193: 639–646.

34. Wohlford-LenaneCL, MeyerholzDK, PerlmanS, ZhouH, TranD, et al. (2009) Rhesus theta-defensin prevents death in a mouse model of severe acute respiratory syndrome coronavirus pulmonary disease. J Virol 83: 11385–11390.

35. Huang daW, ShermanBT, LempickiRA (2009) Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources. Nat Protoc 4: 44–57.

36. FriemanMB, ChenJ, MorrisonTE, WhitmoreA, FunkhouserW, et al. (2010) SARS-CoV pathogenesis is regulated by a STAT1 dependent but a type I, II and III interferon receptor independent mechanism. PLoS Pathog 6: e1000849.

37. SheahanT, MorrisonTE, FunkhouserW, UematsuS, AkiraS, et al. (2008) MyD88 is required for protection from lethal infection with a mouse-adapted SARS-CoV. PLoS Pathog 4: e1000240.

38. FormstecherE, ArestaS, ColluraV, HamburgerA, MeilA, et al. (2005) Protein interaction mapping: a Drosophila case study. Genome Res 15: 376–384.

39. KorollM, RathjenFG, VolkmerH (2001) The neural cell recognition molecule neurofascin interacts with syntenin-1 but not with syntenin-2, both of which reveal self-associating activity. J Biol Chem 276: 10646–10654.

40. YouJ, DoveBK, EnjuanesL, DeDiegoML, AlvarezE, et al. (2005) Subcellular localization of the severe acute respiratory syndrome coronavirus nucleocapsid protein. J Gen Virol 86: 3303–3310.

41. BoukercheH, SuZZ, EmdadL, SarkarD, FisherPB (2007) mda-9/Syntenin regulates the metastatic phenotype in human melanoma cells by activating nuclear factor-kappaB. Cancer Res 67: 1812–1822.

42. KumarS, BoehmJ, LeeJC (2003) p38 MAP kinases: key signalling molecules as therapeutic targets for inflammatory diseases. Nat Rev Drug Discov 2: 717–726.

43. UnderwoodDC, OsbornRR, BochnowiczS, WebbEF, RiemanDJ, et al. (2000) SB 239063, a p38 MAPK inhibitor, reduces neutrophilia, inflammatory cytokines, MMP-9, and fibrosis in lung. Am J Physiol Lung Cell Mol Physiol 279: L895–902.

44. TanYJ, TengE, ShenS, TanTHP, GohPY, et al. (2004) A novel severe acute respiratory syndrome coronavirus protein, U274, is transported to the cell surface and undergoes endocytosis. J Virol 78: 6723–6734.

45. YuanX, LiJ, ShanY, YangZ, ZhaoZ, et al. (2005) Subcellular localization and membrane association of SARS-CoV 3a protein. Virus Res 109: 191–202.

46. EckmannCR, NeunteuflA, PfaffstetterL, JantschMF (2001) The human but not the Xenopus RNA-editing enzyme ADAR1 has an atypical nuclear localization signal and displays the characteristics of a shuttling protein. Mol Biol Cell 12: 1911–1924.

47. CuendaA, RouseJ, DozaYN, MeierR, CohenP, et al. (1995) SB 203580 is a specific inhibitor of a MAP kinase homologue which is stimulated by cellular stresses and interleukin-1. FEBS Lett 364: 229–233.

48. KumarS, JiangMS, AdamsJL, LeeJC (1999) Pyridinylimidazole compound SB 203580 inhibits the activity but not the activation of p38 mitogen-activated protein kinase. Biochem Biophys Res Commun 263: 825–831.

49. GardiolD (2012) PDZ-containing proteins as targets in human pathologies. FEBS J 279: 3529.

50. SubbaiahVK, KranjecC, ThomasM, BanksL (2011) PDZ domains: the building blocks regulating tumorigenesis. Biochem J 439: 195–205.

51. RobertsS, DeluryC, MarshE (2012) The PDZ protein discs-large (DLG): the ‘Jekyll and Hyde’ of the epithelial polarity proteins. FEBS J 279: 3549–3558.

52. NichollsJM, PoonLL, LeeKC, NgWF, LaiST, et al. (2003) Lung pathology of fatal severe acute respiratory syndrome. Lancet 361: 1773–1778.

53. RockxB, BaasT, ZornetzerGA, HaagmansB, SheahanT, et al. (2009) Early upregulation of acute respiratory distress syndrome-associated cytokines promotes lethal disease in an aged-mouse model of severe acute respiratory syndrome coronavirus infection. J Virol 83: 7062–7074.

54. TangNL, ChanPK, WongCK, ToKF, WuAK, et al. (2005) Early enhanced expression of interferon-inducible protein-10 (CXCL-10) and other chemokines predicts adverse outcome in severe acute respiratory syndrome. Clin Chem 51: 2333–2340.

55. GralinskiLE, BankheadA3rd, JengS, MenacheryVD, ProllS, et al. (2013) Mechanisms of severe acute respiratory syndrome coronavirus-induced acute lung injury. MBio 4: e00271–00213.

56. SmitsSL, de LangA, van den BrandJM, LeijtenLM, vanIWF, et al. (2010) Exacerbated innate host response to SARS-CoV in aged non-human primates. PLoS Pathog 6: e1000756.

57. PeirisJS, ChuCM, ChengVC, ChanKS, HungIF, et al. (2003) Clinical progression and viral load in a community outbreak of coronavirus-associated SARS pneumonia: a prospective study. Lancet 361: 1767–1772.

58. LeeCH, ChenRF, LiuJW, YehWT, ChangJC, et al. (2004) Altered p38 mitogen-activated protein kinase expression in different leukocytes with increment of immunosuppressive mediators in patients with severe acute respiratory syndrome. J Immunol 172: 7841–7847.

59. FischerF, StegenCF, MastersPS, SamsonoffWA (1998) Analysis of constructed E gene mutants of mouse hepatitis virus confirms a pivotal role for E protein in coronavirus assembly. J Virol 72: 7885–7894.

60. BorgelingY, SchmolkeM, ViemannD, NordhoffC, RothJ, et al. (2013) Inhibition of p38 Mitogen-activated protein kinase impairs influenza virus-induced primary and secondary host gene responses and protects mice from lethal H5N1 infection. J Biol Chem 45: 13–27.

61. BoukercheH, AissaouiH, PrevostC, HirbecH, DasSK, et al. (2010) Src kinase activation is mandatory for MDA-9/syntenin-mediated activation of nuclear factor-kappaB. Oncogene 29: 3054–3066.

62. AlvarezE, DeDiegoML, Nieto-TorresJL, Jimenez-GuardenoJM, Marcos-VillarL, et al. (2010) The envelope protein of severe acute respiratory syndrome coronavirus interacts with the non-structural protein 3 and is ubiquitinated. Virology 402: 281–291.

63. BustinSA, BenesV, GarsonJA, HellemansJ, HuggettJ, et al. (2009) The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clin Chem 55: 611–622.

64. Fromont-RacineM, RainJC, LegrainP (1997) Toward a functional analysis of the yeast genome through exhaustive two-hybrid screens. Nat Genet 16: 277–282.

65. SmythGK (2004) Linear models and empirical bayes methods for assessing differential expression in microarray experiments. Stat Appl Genet Mol Biol 3: Article3.

66. BenjaminiY, HochbergY (1995) Controlling the false discovery rate: a practical and powerful approach to multiple testing. J Roy Stat Soc B 57: 289–300.

67. ReinerA, YekutieliD, BenjaminiY (2003) Identifying differentially expressed genes using false discovery rate controlling procedures. Bioinformatics 19: 368–375.

68. AlmazanF, DeDiegoML, GalanC, EscorsD, AlvarezE, et al. (2006) Construction of a SARS-CoV infectious cDNA clone and a replicon to study coronavirus RNA synthesis. J Virol 80: 10900–10906.

69. SnijderEJ, BredenbeekPJ, DobbeJC, ThielV, ZiebuhrJ, et al. (2003) Unique and conserved features of genome and proteome of SARS-coronavirus, an early split-off from the coronavirus group 2 lineage. J Mol Biol 331: 991–1004.