The Anti-interferon Activity of Conserved Viral dUTPase ORF54 is Essential for an Effective MHV-68 Infection


Gammaherpesviruses such as KSHV and EBV establish lifelong persistent infections through latency in lymphocytes. These viruses have evolved several strategies to counteract the various components of the innate and adaptive immune systems. We conducted an unbiased screen using the genetically and biologically related virus, MHV-68, to find viral ORFs involved in the inhibition of type I interferon signaling and identified a conserved viral dUTPase, ORF54. Here we define the contribution of ORF54 in type I interferon inhibition by ectopic expression and through the use of genetically modified MHV-68. ORF54 and an ORF54 lacking dUTPase enzymatic activity efficiently inhibit type I interferon signaling by inducing the degradation of the type I interferon receptor protein IFNAR1. Subsequently, we show in vitro that the lack of ORF54 causes a reduction in lytic replication in the presence of type I interferon signaling. Investigation of the physiological consequence of IFNAR1 degradation and importance of ORF54 during MHV-68 in vivo infection demonstrates that ORF54 has an even greater impact on persistent infection than on lytic replication. MHV-68 lacking ORF54 expression is unable to efficiently establish latent infection in lymphocytes, although it replicates relatively normally in lung tissues. However, infection of IFNAR−/− mice alleviates this phenotype, emphasizing the specific role of ORF54 in type I interferon inhibition. Infection of mice and cells by a recombinant MHV-68 virus harboring a site specific mutation in ORF54 rendering the dUTPase inactive demonstrates that dUTPase enzymatic activity is not required for anti-interferon function of ORF54. Moreover, we find that dUTPase activity is dispensable at all stages of MHV-68 infection analyzed. Overall, our data suggest that ORF54 has evolved anti-interferon activity in addition to its dUTPase enzymatic activity, and that it is actually the anti-interferon role that renders ORF54 critical for establishing an effective persistent infection of MHV-68.


Vyšlo v časopise: The Anti-interferon Activity of Conserved Viral dUTPase ORF54 is Essential for an Effective MHV-68 Infection. PLoS Pathog 7(10): e32767. doi:10.1371/journal.ppat.1002292
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1002292

Souhrn

Gammaherpesviruses such as KSHV and EBV establish lifelong persistent infections through latency in lymphocytes. These viruses have evolved several strategies to counteract the various components of the innate and adaptive immune systems. We conducted an unbiased screen using the genetically and biologically related virus, MHV-68, to find viral ORFs involved in the inhibition of type I interferon signaling and identified a conserved viral dUTPase, ORF54. Here we define the contribution of ORF54 in type I interferon inhibition by ectopic expression and through the use of genetically modified MHV-68. ORF54 and an ORF54 lacking dUTPase enzymatic activity efficiently inhibit type I interferon signaling by inducing the degradation of the type I interferon receptor protein IFNAR1. Subsequently, we show in vitro that the lack of ORF54 causes a reduction in lytic replication in the presence of type I interferon signaling. Investigation of the physiological consequence of IFNAR1 degradation and importance of ORF54 during MHV-68 in vivo infection demonstrates that ORF54 has an even greater impact on persistent infection than on lytic replication. MHV-68 lacking ORF54 expression is unable to efficiently establish latent infection in lymphocytes, although it replicates relatively normally in lung tissues. However, infection of IFNAR−/− mice alleviates this phenotype, emphasizing the specific role of ORF54 in type I interferon inhibition. Infection of mice and cells by a recombinant MHV-68 virus harboring a site specific mutation in ORF54 rendering the dUTPase inactive demonstrates that dUTPase enzymatic activity is not required for anti-interferon function of ORF54. Moreover, we find that dUTPase activity is dispensable at all stages of MHV-68 infection analyzed. Overall, our data suggest that ORF54 has evolved anti-interferon activity in addition to its dUTPase enzymatic activity, and that it is actually the anti-interferon role that renders ORF54 critical for establishing an effective persistent infection of MHV-68.


Zdroje

1. KatzeMGHeYGaleMJr 2002 Viruses and interferon: a fight for supremacy. Nat Rev Immunol 2 675 687

2. MossmanKLAshkarAA 2005 Herpesviruses and the innate immune response. Viral Immunol 18 267 281

3. AlamRGorskaM 2003 3. Lymphocytes. J Allergy Clin Immunol 111 S476 485

4. ChaplinDD 2003 1. Overview of the immune response. J Allergy Clin Immunol 111 S442 459

5. TakeuchiOAkiraS 2009 Innate immunity to virus infection. Immunol Rev 227 75 86

6. DurandVWongSYToughDFLe BonA 2004 Shaping of adaptive immune responses to soluble proteins by TLR agonists: a role for IFN-alpha/beta. Immunol Cell Biol 82 596 602

7. LevyDEGarcia-SastreA 2001 The virus battles: IFN induction of the antiviral state and mechanisms of viral evasion. Cytokine Growth Factor Rev 12 143 156

8. SamuelCE 2001 Antiviral actions of interferons. Clin Microbiol Rev 14 778 809, table of contents

9. TaniguchiTTakaokaA 2002 The interferon-alpha/beta system in antiviral responses: a multimodal machinery of gene regulation by the IRF family of transcription factors. Curr Opin Immunol 14 111 116

10. PestkaSLangerJAZoonKCSamuelCE 1987 Interferons and their actions. Annu Rev Biochem 56 727 777

11. de WeerdNASamarajiwaSAHertzogPJ 2007 Type I interferon receptors: biochemistry and biological functions. J Biol Chem 282 20053 20057

12. UzeGLutfallaGMogensenKE 1995 Alpha and beta interferons and their receptor and their friends and relations. J Interferon Cytokine Res 15 3 26

13. CohenBNovickDBarakSRubinsteinM 1995 Ligand-induced association of the type I interferon receptor components. Mol Cell Biol 15 4208 4214

14. RoizmanBaPPE 2001 The Family Herpesviridae: A Brief Introduction. KnipeDMHowleyPMGriffinDEMartinMALambRA Fields Virology. 4 ed Philadelphia Lippincott Williams & Wilkins 2381 2397

15. BishopGABuschLK 2002 Molecular mechanisms of B-lymphocyte transformation by Epstein-Barr virus. Microbes Infect 4 853 857

16. CrawfordDH 2001 Biology and disease associations of Epstein-Barr virus. Philos Trans R Soc Lond B Biol Sci 356 461 473

17. GanemD 1996 Human herpesvirus 8 and the biology of Kaposi's sarcoma. Semin Virol 7 325 332

18. KieffERickinsonAB 2001 Epstein-Barr virus and its replication. KnipeDMHowleyPM Fields Virology Philadelphia Lippincott Williams & Wilkins 2511 2627

19. MoorePChangY 2001 Kaposi's sarcoma-associated herpesvirus. KnipeDMHowleyPM Fields Virology. 4th ed Philadelphia Lippincott Williams & Wilkins 2803 2833

20. MoorePSKingsleyLAHolmbergSDSpiraTGuptaP 1996 Kaposi's sarcoma-associated herpesvirus infection prior to onset of Kaposi's sarcoma. AIDS 10 175 180

21. RoizmanBKnipeDM 2001 Herpes simplex viruses and their replication. KnipeDMHowleyPMGriffinRALambRAMartinMA Fields Virology. fourth ed Philadelphia Lippincott-Raven Publishers 2399 2459

22. SchwarzmannFvon BaehrRJagerMPrangNBohmS 1999 A case of severe chronic active infection with Epstein-Barr virus: immunologic deficiencies associated with a lytic virus strain. Clin Infect Dis 29 626 631

23. YoungLSRickinsonAB 2004 Epstein-Barr virus: 40 years on. Nat Rev Cancer 4 757 768

24. CesarmanEChangYMoorePSSaidJWKnowlesDM 1995 Kaposi's sarcoma-associated herpesvirus-like DNA sequences in AIDS- related body-cavity-based lymphomas. N Engl J Med 332 1186 1191

25. ChangYCesarmanEPessinMSLeeFCulpepperJ 1994 Identification of herpesvirus-like DNA sequences in AIDS-associated Kaposi's sarcoma. Science 266 1865 1869

26. CorbellinoMPoirelLAubinJTPaulliMMagriniU 1996 The role of human herpesvirus 8 and Epstein-Barr virus in the pathogenesis of giant lymph node hyperplasia (Castleman's disease). Clin Infect Dis 22 1120 1121

27. SoulierJGrolletLOksenhendlerECacoubPCazals-HatemD 1995 Kaposi's sarcoma-associated herpesvirus-like DNA sequences in multicentric Castleman's disease. Blood 86 1276 1280

28. Thorley-LawsonDAGrossA 2004 Persistence of the Epstein-Barr virus and the origins of associated lymphomas. N Engl J Med 350 1328 1337

29. Sunil-ChandraNPArnoJFazakerleyJNashAA 1994 Lymphoproliferative disease in mice infected with murine gammaherpesvirus 68. Am J Pathol 145 818 826

30. Sunil-ChandraNPEfstathiouSArnoJNashAA 1992 Virological and pathological features of mice infected with murine gamma-herpesvirus 68. J Gen Virol 73 2347 2356

31. Sunil-ChandraNPEfstathiouSNashAA 1992 Murine gammaherpesvirus 68 establishes a latent infection in mouse B lymphocytes in vivo. J Gen Virol 73 3275 3279

32. VirginHWtLatreillePWamsleyPHallsworthKWeckKE 1997 Complete sequence and genomic analysis of murine gammaherpesvirus 68. J Virol 71 5894 5904

33. SimasJPEfstathiouS 1998 Murine gammaherpesvirus 68: a model for the study of gammaherpesvirus pathogenesis. Trends Microbiol 6 276 282

34. WuTTLiaoHITongLLeangRSSmithG 2011 Construction and characterization of an infectious murine gammaherpesivrus-68 bacterial artificial chromosome. J Biomed Biotechnol 2011 926258

35. BissonSAPageALGanemD 2009 A Kaposi's sarcoma-associated herpesvirus protein that forms inhibitory complexes with type I interferon receptor subunits, Jak and STAT proteins, and blocks interferon-mediated signal transduction. J Virol 83 5056 5066

36. HwangSKimKSFlanoEWuTTTongLM 2009 Conserved herpesviral kinase promotes viral persistence by inhibiting the IRF-3-mediated type I interferon response. Cell Host Microbe 5 166 178

37. SathishNZhuFXGolubEELiangQYuanY 2011 Mechanisms of autoinhibition of IRF-7 and a probable model for inactivation of IRF-7 by Kaposi's sarcoma-associated herpesvirus protein ORF45. J Biol Chem 286 746 756

38. YuYWangSEHaywardGS 2005 The KSHV immediate-early transcription factor RTA encodes ubiquitin E3 ligase activity that targets IRF7 for proteosome-mediated degradation. Immunity 22 59 70

39. CoscoyLGanemD 2000 Kaposi's sarcoma-associated herpesvirus encodes two proteins that block cell surface display of MHC class I chains by enhancing their endocytosis. Proc Natl Acad Sci U S A 97 8051 8056

40. CoscoyLGanemD 2001 A viral protein that selectively downregulates ICAM-1 and B7-2 and modulates T cell costimulation. J Clin Invest 107 1599 1606

41. FruhKBarteeEGouveiaKMansouriM 2002 Immune evasion by a novel family of viral PHD/LAP-finger proteins of gamma-2 herpesviruses and poxviruses. Virus Res 88 55 69

42. MansouriMViswanathanKDouglasJLHinesJGustinJ 2009 Molecular mechanism of BST2/tetherin downregulation by K5/MIR2 of Kaposi's sarcoma-associated herpesvirus. J Virol 83 9672 9681

43. BonameJMStevensonPG 2001 MHC class I ubiquitination by a viral PHD/LAP finger protein. Immunity 15 627 636

44. HarrisJMMcIntoshEMMuscatGE 1999 Structure/function analysis of a dUTPase: catalytic mechanism of a potential chemotherapeutic target. J Mol Biol 288 275 287

45. BaldoAMMcClureMA 1999 Evolution and horizontal transfer of dUTPase-encoding genes in viruses and their hosts. J Virol 73 7710 7721

46. McClureMA 2001 Evolution of the DUT gene: horizontal transfer between host and pathogen in all three domains of life. Curr Protein Pept Sci 2 313 324

47. LiangXShinYCMeansREJungJU 2004 Inhibition of interferon-mediated antiviral activity by murine gammaherpesvirus 68 latency-associated M2 protein. J Virol 78 12416 12427

48. DavisonAJStowND 2005 New genes from old: redeployment of dUTPase by herpesviruses. J Virol 79 12880 12892

49. TarbouriechNBuissonMSeigneurinJMCusackSBurmeisterWP 2005 The monomeric dUTPase from Epstein-Barr virus mimics trimeric dUTPases. Structure 13 1299 1310

50. SongMJHwangSWongWHWuTTLeeS 2005 Identification of viral genes essential for replication of murine gamma-herpesvirus 68 using signature-tagged mutagenesis. Proc Natl Acad Sci U S A 102 3805 3810

51. BrownHJMcBrideWHZackJASunR 2005 Prostratin and bortezomib are novel inducers of latent Kaposi's sarcoma-associated herpesvirus. Antivir Ther 10 745 751

52. DesmyterJMelnickJLRawlsWE 1968 Defectiveness of interferon production and of rubella virus interference in a line of African green monkey kidney cells (Vero). J Virol 2 955 961

53. Weslow-SchmidtJLJewellNAMertzSESimasJPDurbinJE 2007 Type I interferon inhibition and dendritic cell activation during gammaherpesvirus respiratory infection. J Virol 81 9778 9789

54. WuLFossumEJooCHInnKSShinYC 2009 Epstein-Barr virus LF2: an antagonist to type I interferon. J Virol 83 1140 1146

55. RezaeeSACunninghamCDavisonAJBlackbournDJ 2006 Kaposi's sarcoma-associated herpesvirus immune modulation: an overview. J Gen Virol 87 1781 1804

56. GonzalezALarssonGPerssonRCedergren-ZeppezauerE 2001 Atomic resolution structure of Escherichia coli dUTPase determined ab initio. Acta Crystallogr D Biol Crystallogr 57 767 774

57. McGeehanJEDepledgeNWMcGeochDJ 2001 Evolution of the dUTPase gene of mammalian and avian herpesviruses. Curr Protein Pept Sci 2 325 333

58. Martinez-GuzmanDRickabaughTWuTTBrownHColeS 2003 Transcription program of murine gammaherpesvirus 68. J Virol 77 10488 10503

59. LadnerRDMcNultyDECarrSARobertsGDCaradonnaSJ 1996 Characterization of distinct nuclear and mitochondrial forms of human deoxyuridine triphosphate nucleotidohydrolase. J Biol Chem 271 7745 7751

60. CaposioPRieraLHahnGLandolfoSGribaudoG 2004 Evidence that the human cytomegalovirus 46-kDa UL72 protein is not an active dUTPase but a late protein dispensable for replication in fibroblasts. Virology 325 264 276

61. DunnWChouCLiHHaiRPattersonD 2003 Functional profiling of a human cytomegalovirus genome. Proc Natl Acad Sci U S A 100 14223 14228

62. AbateDAWatanabeSMocarskiES 2004 Major human cytomegalovirus structural protein pp65 (ppUL83) prevents interferon response factor 3 activation in the interferon response. J Virol 78 10995 11006

63. BrowneEPShenkT 2003 Human cytomegalovirus UL83-coded pp65 virion protein inhibits antiviral gene expression in infected cells. Proc Natl Acad Sci U S A 100 11439 11444

64. WaldmanWJWilliamsMVJrLemeshowSBinkleyPGuttridgeD 2008 Epstein-Barr virus-encoded dUTPase enhances proinflammatory cytokine production by macrophages in contact with endothelial cells: evidence for depression-induced atherosclerotic risk. Brain Behav Immun 22 215 223

65. PadgettDAHotchkissAKPyterLMNelsonRJYangE 2004 Epstein-Barr virus-encoded dUTPase modulates immune function and induces sickness behavior in mice. J Med Virol 74 442 448

66. ArizaMEGlaserRKaumayaPTJonesCWilliamsMV 2009 The EBV-encoded dUTPase activates NF-kappa B through the TLR2 and MyD88-dependent signaling pathway. J Immunol 182 851 859

67. GlaserRLitskyMLPadgettDABaiocchiRAYangEV 2006 EBV-encoded dUTPase induces immune dysregulation: Implications for the pathophysiology of EBV-associated disease. Virology 346 205 218

68. WilliamsMV 1984 Deoxyuridine triphosphate nucleotidohydrolase induced by herpes simplex virus type 1. Purification and characterization of induced enzyme. J Biol Chem 259 10080 10084

69. WilliamsMVParrisDS 1987 Characterization of a herpes simplex virus type 2 deoxyuridine triphosphate nucleotidohydrolase and mapping of a gene conferring type specificity for the enzyme. Virology 156 282 292

70. BarkerDERoizmanB 1990 Identification of three genes nonessential for growth in cell culture near the right terminus of the unique sequences of long component of herpes simplex virus 1. Virology 177 684 691

71. FisherFBPrestonVG 1986 Isolation and characterisation of herpes simplex virus type 1 mutants which fail to induce dUTPase activity. Virology 148 190 197

72. ColemanHMde LimaBMortonVStevensonPG 2003 Murine gammaherpesvirus 68 lacking thymidine kinase shows severe attenuation of lytic cycle replication in vivo but still establishes latency. J Virol 77 2410 2417

73. GillMBMayJSColacoSStevensonPG 2010 Important role for the murid herpesvirus 4 ribonucleotide reductase large subunit in host colonization via the respiratory tract. J Virol 84 10937 10942

74. PylesRBSawtellNMThompsonRL 1992 Herpes simplex virus type 1 dUTPase mutants are attenuated for neurovirulence, neuroinvasiveness, and reactivation from latency. J Virol 66 6706 6713

75. WuTTBlackmanMASunR 2010 Prospects of a novel vaccination strategy for human gamma-herpesviruses. Immunol Res 48 122 146

76. CaradonnaSJAdamkiewiczDM 1984 Purification and properties of the deoxyuridine triphosphate nucleotidohydrolase enzyme derived from HeLa S3 cells. Comparison to a distinct dUTP nucleotidohydrolase induced in herpes simplex virus-infected HeLa S3 cells. J Biol Chem 259 5459 5464

77. SmithGAEnquistLW 1999 Construction and transposon mutagenesis in Escherichia coli of a full-length infectious clone of pseudorabies virus, an alphaherpesvirus. J Virol 73 6405 6414

78. SmithGAEnquistLW 2000 A self-recombining bacterial artificial chromosome and its application for analysis of herpesvirus pathogenesis. Proc Natl Acad Sci U S A 97 4873 4878

Štítky
Hygiena a epidemiológia Infekčné lekárstvo Laboratórium

Článok vyšiel v časopise

PLOS Pathogens


2011 Číslo 10
Najčítanejšie tento týždeň
Najčítanejšie v tomto čísle
Kurzy

Zvýšte si kvalifikáciu online z pohodlia domova

Eozinofilní granulomatóza s polyangiitidou
nový kurz
Autori: doc. MUDr. Martina Doubková, Ph.D.

Všetky kurzy
Prihlásenie
Zabudnuté heslo

Zadajte e-mailovú adresu, s ktorou ste vytvárali účet. Budú Vám na ňu zasielané informácie k nastaveniu nového hesla.

Prihlásenie

Nemáte účet?  Registrujte sa