The SARS-Coronavirus-Host Interactome: Identification of Cyclophilins as Target for Pan-Coronavirus Inhibitors


Coronaviruses (CoVs) are important human and animal pathogens that induce fatal respiratory, gastrointestinal and neurological disease. The outbreak of the severe acute respiratory syndrome (SARS) in 2002/2003 has demonstrated human vulnerability to (Coronavirus) CoV epidemics. Neither vaccines nor therapeutics are available against human and animal CoVs. Knowledge of host cell proteins that take part in pivotal virus-host interactions could define broad-spectrum antiviral targets. In this study, we used a systems biology approach employing a genome-wide yeast-two hybrid interaction screen to identify immunopilins (PPIA, PPIB, PPIH, PPIG, FKBP1A, FKBP1B) as interaction partners of the CoV non-structural protein 1 (Nsp1). These molecules modulate the Calcineurin/NFAT pathway that plays an important role in immune cell activation. Overexpression of NSP1 and infection with live SARS-CoV strongly increased signalling through the Calcineurin/NFAT pathway and enhanced the induction of interleukin 2, compatible with late-stage immunopathogenicity and long-term cytokine dysregulation as observed in severe SARS cases. Conversely, inhibition of cyclophilins by cyclosporine A (CspA) blocked the replication of CoVs of all genera, including SARS-CoV, human CoV-229E and -NL-63, feline CoV, as well as avian infectious bronchitis virus. Non-immunosuppressive derivatives of CspA might serve as broad-range CoV inhibitors applicable against emerging CoVs as well as ubiquitous pathogens of humans and livestock.


Vyšlo v časopise: The SARS-Coronavirus-Host Interactome: Identification of Cyclophilins as Target for Pan-Coronavirus Inhibitors. PLoS Pathog 7(10): e32767. doi:10.1371/journal.ppat.1002331
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1002331

Souhrn

Coronaviruses (CoVs) are important human and animal pathogens that induce fatal respiratory, gastrointestinal and neurological disease. The outbreak of the severe acute respiratory syndrome (SARS) in 2002/2003 has demonstrated human vulnerability to (Coronavirus) CoV epidemics. Neither vaccines nor therapeutics are available against human and animal CoVs. Knowledge of host cell proteins that take part in pivotal virus-host interactions could define broad-spectrum antiviral targets. In this study, we used a systems biology approach employing a genome-wide yeast-two hybrid interaction screen to identify immunopilins (PPIA, PPIB, PPIH, PPIG, FKBP1A, FKBP1B) as interaction partners of the CoV non-structural protein 1 (Nsp1). These molecules modulate the Calcineurin/NFAT pathway that plays an important role in immune cell activation. Overexpression of NSP1 and infection with live SARS-CoV strongly increased signalling through the Calcineurin/NFAT pathway and enhanced the induction of interleukin 2, compatible with late-stage immunopathogenicity and long-term cytokine dysregulation as observed in severe SARS cases. Conversely, inhibition of cyclophilins by cyclosporine A (CspA) blocked the replication of CoVs of all genera, including SARS-CoV, human CoV-229E and -NL-63, feline CoV, as well as avian infectious bronchitis virus. Non-immunosuppressive derivatives of CspA might serve as broad-range CoV inhibitors applicable against emerging CoVs as well as ubiquitous pathogens of humans and livestock.


Zdroje

1. PerlmanSNetlandJ 2009 Coronaviruses post-SARS: update on replication and pathogenesis. Nat Rev Microbiol 7 439 450

2. PeirisJSYuenKYOsterhausADStohrK 2003 The severe acute respiratory syndrome. N Engl J Med 349 2431 2441

3. WangLFShiZZhangSFieldHDaszakP 2006 Review of bats and SARS. Emerg Infect Dis 12 1834 1840

4. DrexlerJFGloza-RauschFGlendeJCormanVMMuthD 2010 Genomic characterization of severe acute respiratory syndrome-related coronavirus in European bats and classification of coronaviruses based on partial RNA-dependent RNA polymerase gene sequences. J Virol 84 11336 11349

5. WooPCLauSKHuangYYuenKY 2009 Coronavirus diversity, phylogeny and interspecies jumping. Exp Biol Med (Maywood) 234 1117 1127

6. von BrunnATeepeCSimpsonJCPepperkokRFriedelCC 2007 Analysis of intraviral protein-protein interactions of the SARS coronavirus ORFeome. PLoS One 2 e459

7. UetzPDongYAZeretzkeCAtzlerCBaikerA 2006 Herpesviral protein networks and their interaction with the human proteome. Science 311 239 242

8. FeskeSGiltnaneJDolmetschRStaudtLMRaoA 2001 Gene regulation mediated by calcium signals in T lymphocytes. Nat Immunol 2 316 324

9. HoganPGChenLNardoneJRaoA 2003 Transcriptional regulation by calcium, calcineurin, and NFAT. Genes Dev 17 2205 2232

10. BraunPTasanMDrezeMBarrios-RodilesMLemmensI 2009 An experimentally derived confidence score for binary protein-protein interactions. Nat Methods 6 91 97

11. AlexaARahnenfuhrerJLengauerT 2006 Improved scoring of functional groups from gene expression data by decorrelating GO graph structure 3. Bioinformatics 22 1600 1607

12. BauerSGrossmannSVingronMRobinsonPN 2008 Ontologizer 2.0—a multifunctional tool for GO term enrichment analysis and data exploration. Bioinformatics 24 1650 1651

13. YuHKimPMSprecherETrifonovVGersteinM 2007 The importance of bottlenecks in protein networks: correlation with gene essentiality and expression dynamics. PLoS Comput Biol 3 e59

14. Hermann-KleiterNBaierG 2010 NFAT pulls the strings during CD4+ T helper cell effector functions. Blood 115 2989 2997

15. LubanJBossoltKLFrankeEKKalpanaGVGoffSP 1993 Human immunodeficiency virus type 1 Gag protein binds to cyclophilins A and B. Cell 73 1067 1078

16. WatashiKIshiiNHijikataMInoueDMurataT 2005 Cyclophilin B is a functional regulator of hepatitis C virus RNA polymerase. Mol Cell 19 111 122

17. MacianF 2005 NFAT proteins: key regulators of T-cell development and function 7. Nat Rev Immunol 5 472 484

18. PfefferleSKraehlingVDittVGrywnaKMuehlbergerE 2009 Reverse genetic characterization of the natural genomic deletion in SARS-Coronavirus strain Frankfurt-1 open reading frame 7b reveals an attenuating function of the 7b protein in-vitro and in-vivo. Virol J 6 131

19. SpiegelMWeberF 2006 Inhibition of cytokine gene expression and induction of chemokine genes in non-lymphatic cells infected with SARS coronavirus. Virol J 3 17

20. JainJLohCRaoA 1995 Transcriptional regulation of the IL-2 gene. Curr Opin Immunol 7 333 342

21. YoneyamaMSuharaWFukuharaYFukudaMNishidaE 1998 Direct triggering of the type I interferon system by virus infection: activation of a transcription factor complex containing IRF-3 and CBP/p300. EMBO J 17 1087 1095

22. WangYShiHRigoletPWuNZhuL 2010 Nsp1 proteins of group I and SARS coronaviruses share structural and functional similarities. Infect Genet Evol 10 919 924

23. ChenLGuiCLuoXYangQGuntherS 2005 Cinanserin is an inhibitor of the 3C-like proteinase of severe acute respiratory syndrome coronavirus and strongly reduces virus replication in vitro. J Virol 79 7095 7103

24. AlmazanFDediegoMLGalanCEscorsDAlvarezE 2006 Construction of a severe acute respiratory syndrome coronavirus infectious cDNA clone and a replicon to study coronavirus RNA synthesis. J Virol 80 10900 10906

25. YountBCurtisKMFritzEAHensleyLEJahrlingPB 2003 Reverse genetics with a full-length infectious cDNA of severe acute respiratory syndrome coronavirus. Proc Natl Acad Sci U S A 100 12995 13000

26. ErikssonKKMakiaDThielV 2008 Generation of recombinant coronaviruses using vaccinia virus as the cloning vector and stable cell lines containing coronaviral replicon RNAs. Methods Mol Biol 454 237 254

27. PanJPengXGaoYLiZLuX 2008 Genome-wide analysis of protein-protein interactions and involvement of viral proteins in SARS-CoV replication. PLoS One 3 e3299

28. ImbertISnijderEJDimitrovaMGuillemotJCLecineP 2008 The SARS-Coronavirus PLnc domain of nsp3 as a replication/transcription scaffolding protein. Virus Res 133 136 148

29. NgLFHibberdMLOoiEETangKFNeoSY 2004 A human in vitro model system for investigating genome-wide host responses to SARS coronavirus infection. BMC Infect Dis 4 34

30. PangRTPoonTCChanKCLeeNLChiuRW 2006 Serum proteomic fingerprints of adult patients with severe acute respiratory syndrome. Clin Chem 52 421 429

31. NeumanBWJosephJSSaikatenduKSSerranoPChatterjeeA 2008 Proteomics analysis unravels the functional repertoire of coronavirus nonstructural protein 3 2. J Virol 82 5279 5294

32. WalhoutAJVidalM 2001 High-throughput yeast two-hybrid assays for large-scale protein interaction mapping. Methods 24 297 306

33. WalhoutAJTempleGFBraschMAHartleyJLLorsonMA 2000 GATEWAY recombinational cloning: application to the cloning of large numbers of open reading frames or ORFeomes. Methods Enzymol 328 575 592

34. Barrios-RodilesMBrownKROzdamarBBoseRLiuZ 2005 High-throughput mapping of a dynamic signaling network in mammalian cells. Science 307 1621 1625

35. ZustRCervantes-BarraganLKuriTBlakqoriGWeberF 2007 Coronavirus non-structural protein 1 is a major pathogenicity factor: implications for the rational design of coronavirus vaccines. PLoS Pathog 3 e109

36. KamitaniWHuangCNarayananKLokugamageKGMakinoS 2009 A two-pronged strategy to suppress host protein synthesis by SARS coronavirus Nsp1 protein. Nat Struct Mol Biol 16 1134 1140

37. WatheletMGOrrMFriemanMBBaricRS 2007 Severe acute respiratory syndrome coronavirus evades antiviral signaling: role of nsp1 and rational design of an attenuated strain. J Virol 81 11620 11633

38. NarayananKHuangCLokugamageKKamitaniWIkegamiT 2008 Severe acute respiratory syndrome coronavirus nsp1 suppresses host gene expression, including that of type I interferon, in infected cells. J Virol 82 4471 4479

39. JonesBMMaESPeirisJSWongPCHoJC 2004 Prolonged disturbances of in vitro cytokine production in patients with severe acute respiratory syndrome (SARS) treated with ribavirin and steroids. Clin Exp Immunol 135 467 473

40. LoutfyMRBlattLMSiminovitchKAWardSWolffB 2003 Interferon alfacon-1 plus corticosteroids in severe acute respiratory syndrome: a preliminary study. JAMA 290 3222 3228

41. PeirisJSChuCMChengVCChanKSHungIF 2003 Clinical progression and viral load in a community outbreak of coronavirus-associated SARS pneumonia: a prospective study. Lancet 361 1767 1772

42. LiuJFarmerJDJrLaneWSFriedmanJWeissmanI 1991 Calcineurin is a common target of cyclophilin-cyclosporin A and FKBP-FK506 complexes. Cell 66 807 815

43. GalatABuaJ 2010 Molecular aspects of cyclophilins mediating therapeutic actions of their ligands. Cell Mol Life Sci 67 3467 3488

44. LuoCLuoHZhengSGuiCYueL 2004 Nucleocapsid protein of SARS coronavirus tightly binds to human cyclophilin A. Biochem Biophys Res Commun 321 557 565

45. FischerGGallayPHopkinsS 2010 Cyclophilin inhibitors for the treatment of HCV infection. Curr Opin Investig Drugs 11 911 918

46. BrockwaySMDenisonMR 2005 Mutagenesis of the murine hepatitis virus nsp1-coding region identifies residues important for protein processing, viral RNA synthesis, and viral replication. Virology 340 209 223

47. AlbersMKranzHKoberIKaiserCKlinkM 2005 Automated yeast two-hybrid screening for nuclear receptor-interacting proteins. Mol Cell Proteomics 4 205 213

48. DrostenCPreiserWGuntherSSchmitzHDoerrHW 2003 Severe acute respiratory syndrome: identification of the etiological agent. Trends Mol Med 9 325 327

49. HerzogPDrostenCMullerMA 2008 Plaque assay for human coronavirus NL63 using human colon carcinoma cells. Virol J 5 138

50. KremplCBallesterosMLZimmerGEnjuanesLKlenkHD 2000 Characterization of the sialic acid binding activity of transmissible gastroenteritis coronavirus by analysis of haemagglutination-deficient mutants. J GenVirol 81 489 496

51. PfefferleSKrahlingVDittVGrywnaKMuhlbergerE 2009 Reverse genetic characterization of the natural genomic deletion in SARS-Coronavirus strain Frankfurt-1 open reading frame 7b reveals an attenuating function of the 7b protein in-vitro and in-vivo. Virol J 6 131

52. BenjaminiYHochbergY 1995 Controlling the false discovery rate: a practical and powerful approach to multiple testing. J R Stat Soc Ser B Stat Methodol 57 289 300

53. RueppABraunerBDunger-KaltenbachIFrishmanGMontroneC 2008 CORUM: the comprehensive resource of mammalian protein complexes 1. Nucleic Acids Res 36 D646 D650

54. PeriSNavarroJDKristiansenTZAmanchyRSurendranathV 2004 Human protein reference database as a discovery resource for proteomics. Nucleic Acids Res 32 D497 D501

55. BreitkreutzBJStarkCRegulyTBoucherLBreitkreutzA 2008 The BioGRID Interaction Database: 2008 update. Nucleic Acids Res 36 D637 D640

56. Team RDC 2007 R: a language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria

57. MarraMAJonesSJAstellCRHoltRABrooks-WilsonA 2003 The Genome sequence of the SARS-associated coronavirus. Science 300 1399 1404

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Hygiena a epidemiológia Infekčné lekárstvo Laboratórium

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