The Apoptogenic Toxin AIP56 Is a Metalloprotease A-B Toxin that Cleaves NF-κb P65

English version České info

AIP56 (apoptosis-inducing protein of 56 kDa) is a major virulence factor of Photobacterium damselae piscicida (Phdp), a Gram-negative pathogen that causes septicemic infections, which are among the most threatening diseases in mariculture. The toxin triggers apoptosis of host macrophages and neutrophils through a process that, in vivo, culminates with secondary necrosis of the apoptotic cells contributing to the necrotic lesions observed in the diseased animals. Here, we show that AIP56 is a NF-κB p65-cleaving zinc-metalloprotease whose catalytic activity is required for the apoptogenic effect. Most of the bacterial effectors known to target NF-κB are type III secreted effectors. In contrast, we demonstrate that AIP56 is an A-B toxin capable of acting at distance, without requiring contact of the bacteria with the target cell. We also show that the N-terminal domain cleaves NF-κB at the Cys³⁹-Glu⁴⁰ peptide bond and that the C-terminal domain is involved in binding and internalization into the cytosol.

Vyšlo v časopise: The Apoptogenic Toxin AIP56 Is a Metalloprotease A-B Toxin that Cleaves NF-κb P65. PLoS Pathog 9(2): e32767. doi:10.1371/journal.ppat.1003128
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1003128

Souhrn

Zdroje

1. GilmoreTD, WolenskiFS (2012) NF-κB: where did it come from and why? Immunol Rev 246: 14–35.

2. RahmanMM, McFaddenG (2011) Modulation of NF-kappaB signalling by microbial pathogens. Nat Rev Microbiol 9: 291–306.

3. Le NegrateG (2012) Subversion of innate immune responses by bacterial hindrance of NF-kappaB pathway. Cell Microbiol 14: 155–167.

4. NeishAS, NaumannM (2011) Microbial-induced immunomodulation by targeting the NF-kappaB system. Trends Microbiol 19: 596–605.

5. MagariñosB, ToranzoAE, RomaldeJL (1996) Phenotypic and pathobiological characteristics of Pasteurella piscicida. Annu Rev Fish Dis 6: 41–64.

6. RomaldeJL (2002) Photobacterium damselae subsp. piscicida: an integrated view of a bacterial fish pathogen. Int Microbiol 5: 3–9.

7. ThuneRL, StanleyLA, CooperRK (1993) Pathogenesis of Gram-negative bacterial infections in warmwater fish. Annu Rev Fish Dis 3: 37–68.

8. BarnesAC, dos SantosNM, EllisAE (2005) Update on bacterial vaccines: Photobacterium damselae subsp. piscicida. Dev Biol (Basel) 121: 75–84.

9. HawkeJP, PlakasSM, Vernon MintonR, McPhearsonRM, SniderTG, et al. (1987) Fish pasteurellosis of cultured striped bass (Morone saxatilis) in coastal Alabama. Aquaculture 65: 193–204.

10. ToranzoAE, BarreiroS, CasalJF, FiguerasA, MagarinosB, et al. (1991) Pasteurellosis in cultured gilthead seabream (Sparus aurata): first report in Spain. Aquaculture 99: 1–15.

11. MagariñosB, SantosY, RomaldeJL, RivasC, BarjaJL, et al. (1992) Pathogenic activities of live cells and extracellular products of the fish pathogen Pasteurella piscicida. J Gen Microbiol 138: 2491–2498.

12. KusudaR, SalatiF (1993) Major bacterial diseases affecting mariculture in Japan. Annu Rev Fish Dis 3: 69–85.

13. NoyaM, MagarinosB, ToranzoAE, LamasJ (1995) Sequential pathology of experimental pasteurellosis in gilthead seabream Sparus aurata. A light- and electron-microscopic study. Dis Aquat Org 21: 177–186.

14. BakopoulosV, PericZ, RodgerH, AdamsA, RichardsR (1997) First report of fish pasteurellosis from Malta. J Aquat Anim Health 9: 26–33.

15. PoulosC, BakopoulosV, ZolotaV, dimitriadisGJ (2004) Histopathological findings after sea bass (Dicentrarchus labrax L.) exposure to extracellular products of Photobacterium damselae subsp. piscicida produced in vivo. Aquac Res 35: 931–936.

16. BakopoulosV, HanifA, PoulosK, GaleottiM, AdamsA, et al. (2004) The effect of in vivo growth on the cellular and extracellular components of the marine bacterial pathogen Photobacterium damsela subsp. piscicida. J Fish Dis 27: 1–13.

17. do ValeA, Costa-RamosC, SilvaA, SilvaDS, GartnerF, et al. (2007) Systemic macrophage and neutrophil destruction by secondary necrosis induced by a bacterial exotoxin in a Gram-negative septicaemia. Cell Microbiol 9: 988–1003.

18. do ValeA, MarquesF, SilvaMT (2003) Apoptosis of sea bass (Dicentrarchus labrax L.) neutrophils and macrophages induced by experimental infection with Photobacterium damselae subsp. piscicida. Fish Shellfish Immunol 15: 129–144.

19. Costa-RamosC, ValeAD, LudovicoP, Dos SantosNM, SilvaMT (2011) The bacterial exotoxin AIP56 induces fish macrophage and neutrophil apoptosis using mechanisms of the extrinsic and intrinsic pathways. Fish Shellfish Immunol 30: 173–181.

20. SilvaMT, do ValeA, dos SantosNM (2008) Secondary necrosis in multicellular animals: an outcome of apoptosis with pathogenic implications. Apoptosis 13: 463–482.

21. do ValeA, SilvaMT, dos SantosNM, NascimentoDS, Reis-RodriguesP, et al. (2005) AIP56, a novel plasmid-encoded virulence factor of Photobacterium damselae subsp. piscicida with apoptogenic activity against sea bass macrophages and neutrophils. Mol Microbiol 58: 1025–1038.

22. SchiavoG, PoulainB, RossettoO, BenfenatiF, TaucL, et al. (1992) Tetanus toxin is a zinc protein and its inhibition of neurotransmitter release and protease activity depend on zinc. EMBO J 11: 3577–3583.

23. SilvaMT, Dos SantosNM, do ValeA (2010) AIP56: A Novel Bacterial Apoptogenic Toxin. Toxins (Basel) 2: 905–918.

24. DegnanPH, YuY, SisnerosN, WingRA, MoranNA (2009) Hamiltonella defensa, genome evolution of protective bacterial endosymbiont from pathogenic ancestors. Proc Natl Acad Sci U S A 106: 9063–9068.

25. CollierRJ (2001) Understanding the mode of action of diphtheria toxin: a perspective on progress during the 20th century. Toxicon 39: 1793–1803.

26. SchiavoG, MatteoliM, MontecuccoC (2000) Neurotoxins affecting neuroexocytosis. Physiol Rev 80: 717–766.

27. BaruchK, Gur-ArieL, NadlerC, KobyS, YerushalmiG, et al. (2010) Metalloprotease type III effectors that specifically cleave JNK and NF-kappaB. EMBO J 30: 221–231.

28. MuhlenS, Ruchaud-SparaganoMH, KennyB (2011) Proteasome-independent degradation of canonical NFkappaB complex components by the NleC protein of pathogenic Escherichia coli. J Biol Chem 286: 5100–5107.

29. PearsonJS, RiedmaierP, MarchesO, FrankelG, HartlandEL (2011) A type III effector protease NleC from enteropathogenic Escherichia coli targets NF-kappaB for degradation. Mol Microbiol 80: 219–230.

30. YenH, OokaT, IguchiA, HayashiT, SugimotoN, et al. (2010) NleC, a type III secretion protease, compromises NF-kappaB activation by targeting p65/RelA. PLoS Pathog 6: e1001231.

31. ShamHP, ShamesSR, CroxenMA, MaC, ChanJM, et al. (2011) Attaching and effacing bacterial effector NleC suppresses epithelial inflammatory responses by inhibiting NF-kappaB and p38 mitogen-activated protein kinase activation. Infect Immun 79: 3552–3562.

32. CoirasM, Lopez-HuertasMR, MateosE, AlcamiJ (2008) Caspase-3-mediated cleavage of p65/RelA results in a carboxy-terminal fragment that inhibits IkappaBalpha and enhances HIV-1 replication in human T lymphocytes. Retrovirology 5: 109.

33. KangKH, LeeKH, KimMY, ChoiKH (2001) Caspase-3-mediated cleavage of the NF-kappa B subunit p65 at the NH2 terminus potentiates naphthoquinone analog-induced apoptosis. J Biol Chem 276: 24638–24644.

34. CollierRJ (2009) Membrane translocation by anthrax toxin. Mol Aspects Med 30: 413–422.

35. SandvigK, OlsnesS (1981) Rapid entry of nicked diphtheria toxin into cells at low pH. Characterization of the entry process and effects of low pH on the toxin molecule. J Biol Chem 256: 9068–9076.

36. SchiavoG, PapiniE, GennaG, MontecuccoC (1990) An intact interchain disulfide bond is required for the neurotoxicity of tetanus toxin. Infect Immun 58: 4136–4141.

37. de PaivaA, AshtonAC, ForanP, SchiavoG, MontecuccoC, et al. (1993) Botulinum A like type B and tetanus toxins fulfils criteria for being a zinc-dependent protease. J Neurochem 61: 2338–2341.

38. BegAA, BaltimoreD (1996) An essential role for NF-kappaB in preventing TNF-alpha-induced cell death. Science 274: 782–784.

39. BegAA, ShaWC, BronsonRT, GhoshS, BaltimoreD (1995) Embryonic lethality and liver degeneration in mice lacking the RelA component of NF-kappa B. Nature 376: 167–170.

40. SenN, Paul BinduD, Gadalla MoatazM, Mustafa AsifK, SenT, et al. (2012) Hydrogen Sulfide-Linked Sulfhydration of NF-κB Mediates Its Antiapoptotic Actions. Mol Cell 45: 13–24.

41. LamkanfiM, DixitVM (2010) Manipulation of host cell death pathways during microbial infections. Cell Host Microbe 8: 44–54.

42. DeLeoFR (2004) Modulation of phagocyte apoptosis by bacterial pathogens. Apoptosis 9: 399–413.

43. WeinrauchY, ZychlinskyA (1999) The induction of apoptosis by bacterial pathogens. Annu Rev Microbiol 53: 155–187.

44. KlimpelKR, AroraN, LepplaSH (1994) Anthrax toxin lethal factor contains a zinc metalloprotease consensus sequence which is required for lethal toxin activity. Mol Microbiol 13: 1093–1100.

45. SchiavoG, RossettoO, BenfenatiF, PoulainB, MontecuccoC (1994) Tetanus and botulinum neurotoxins are zinc proteases specific for components of the neuroexocytosis apparatus. Ann N Y Acad Sci 710: 65–75.

46. ChenFE, HuangDB, ChenYQ, GhoshG (1998) Crystal structure of p50/p65 heterodimer of transcription factor NF-kappaB bound to DNA. Nature 391: 410–413.

47. ChenYQ, GhoshS, GhoshG (1998) A novel DNA recognition mode by the NF-kappa B p65 homodimer. Nat Struct Biol 5: 67–73.

48. HoffmannA, NatoliG, GhoshG (2006) Transcriptional regulation via the NF-kappaB signaling module. Oncogene 25: 6706–6716.

49. KelleherZT, MatsumotoA, StamlerJS, MarshallHE (2007) NOS2 regulation of NF-kappaB by S-nitrosylation of p65. J Biol Chem 282: 30667–30672.

50. Garcia-PineresAJ, CastroV, MoraG, SchmidtTJ, StrunckE, et al. (2001) Cysteine 38 in p65/NF-kappaB plays a crucial role in DNA binding inhibition by sesquiterpene lactones. J Biol Chem 276: 39713–39720.

51. Garcia-PineresAJ, LindenmeyerMT, MerfortI (2004) Role of cysteine residues of p65/NF-kappaB on the inhibition by the sesquiterpene lactone parthenolide and N-ethyl maleimide, and on its transactivating potential. Life Sci 75: 841–856.

52. HanY, EnglertJA, YangR, DeludeRL, FinkMP (2005) Ethyl pyruvate inhibits nuclear factor-kappaB-dependent signaling by directly targeting p65. J Pharmacol Exp Ther 312: 1097–1105.

53. SethiG, AhnKS, AggarwalBB (2008) Targeting nuclear factor-kappa B activation pathway by thymoquinone: role in suppression of antiapoptotic gene products and enhancement of apoptosis. Mol Cancer Res 6: 1059–1070.

54. WatanabeM, NakashimaM, ToganoT, HigashiharaM, WatanabeT, et al. (2008) Identification of the RelA domain responsible for action of a new NF-kappaB inhibitor DHMEQ. Biochem Biophys Res Commun 376: 310–314.

55. LagaM, CottynA, Van HerrewegheF, Vanden BergheW, HaegemanG, et al. (2007) Methylglyoxal suppresses TNF-alpha-induced NF-kappaB activation by inhibiting NF-kappaB DNA-binding. Biochem Pharmacol 74: 579–589.

56. HaKH, ByunMS, ChoiJ, JeongJ, LeeKJ, et al. (2009) N-tosyl-L-phenylalanine chloromethyl ketone inhibits NF-kappaB activation by blocking specific cysteine residues of IkappaB kinase beta and p65/RelA. Biochemistry 48: 7271–7278.

57. HarikumarKB, KunnumakkaraAB, AhnKS, AnandP, KrishnanS, et al. (2009) Modification of the cysteine residues in IkappaBalpha kinase and NF-kappaB (p65) by xanthohumol leads to suppression of NF-kappaB-regulated gene products and potentiation of apoptosis in leukemia cells. Blood 113: 2003–2013.

58. LiangMC, BardhanS, PaceEA, RosmanD, BeutlerJA, et al. (2006) Inhibition of transcription factor NF-kappaB signaling proteins IKKbeta and p65 through specific cysteine residues by epoxyquinone A monomer: correlation with its anti-cancer cell growth activity. Biochem Pharmacol 71: 634–645.

59. SandurSK, IchikawaH, SethiG, AhnKS, AggarwalBB (2006) Plumbagin (5-hydroxy-2-methyl-1,4-naphthoquinone) suppresses NF-kappaB activation and NF-kappaB-regulated gene products through modulation of p65 and IkappaBalpha kinase activation, leading to potentiation of apoptosis induced by cytokine and chemotherapeutic agents. J Biol Chem 281: 17023–17033.

60. StrausDS, PascualG, LiM, WelchJS, RicoteM, et al. (2000) 15-deoxy-delta 12,14-prostaglandin J2 inhibits multiple steps in the NF-kappa B signaling pathway. Proc Natl Acad Sci U S A 97: 4844–4849.

61. WangCY, MayoMW, BaldwinASJr (1996) TNF- and cancer therapy-induced apoptosis: potentiation by inhibition of NF-kappaB. Science 274: 784–787.

62. HaaseR, KirschningCJ, SingA, SchrottnerP, FukaseK, et al. (2003) A dominant role of Toll-like receptor 4 in the signaling of apoptosis in bacteria-faced macrophages. J Immunol 171: 4294–4303.

63. HsuLC, ParkJM, ZhangK, LuoJL, MaedaS, et al. (2004) The protein kinase PKR is required for macrophage apoptosis after activation of Toll-like receptor 4. Nature 428: 341–345.

64. RuckdeschelK, HarbS, RoggenkampA, HornefM, ZumbihlR, et al. (1998) Yersinia enterocolitica impairs activation of transcription factor NF-kappaB: involvement in the induction of programmed cell death and in the suppression of the macrophage tumor necrosis factor alpha production. J Exp Med 187: 1069–1079.

65. MonackDM, MecsasJ, BouleyD, FalkowS (1998) Yersinia-induced apoptosis in vivo aids in the establishment of a systemic infection of mice. J Exp Med 188: 2127–2137.

66. RuckdeschelK, MannelO, RichterK, JacobiCA, TrulzschK, et al. (2001) Yersinia outer protein P of Yersinia enterocolitica simultaneously blocks the nuclear factor-kappa B pathway and exploits lipopolysaccharide signaling to trigger apoptosis in macrophages. J Immunol 166: 1823–1831.

67. JonesRM, LuoL, MobergKH (2012) Aeromonas salmonicida-secreted protein AopP is a potent inducer of apoptosis in a mammalian and a Drosophila model. Cell Microbiol 14: 274–285.

68. ZhangY, TingAT, MarcuKB, BliskaJB (2005) Inhibition of MAPK and NF-κB Pathways Is Necessary for Rapid Apoptosis in Macrophages Infected with Yersinia. J Immunol 174: 7939–7949.

69. ZhouH, MonackDM, KayagakiN, WertzI, YinJ, et al. (2005) Yersinia virulence factor YopJ acts as a deubiquitinase to inhibit NF-kappa B activation. J Exp Med 202: 1327–1332.

70. MukherjeeS, KeitanyG, LiY, WangY, BallHL, et al. (2006) Yersinia YopJ acetylates and inhibits kinase activation by blocking phosphorylation. Science 312: 1211–1214.

71. FehrD, CasanovaC, LivermanA, BlazkovaH, OrthK, et al. (2006) AopP, a type III effector protein of Aeromonas salmonicida, inhibits the NF-kappaB signalling pathway. Microbiology 152: 2809–2818.

72. BhattacharjeeRN, ParkKS, KumagaiY, OkadaK, YamamotoM, et al. (2006) VP1686, a Vibrio type III secretion protein, induces toll-like receptor-independent apoptosis in macrophage through NF-kappaB inhibition. J Biol Chem 281: 36897–36904.

73. MontecuccoC, PapiniE, SchiavoG (1994) Bacterial protein toxins penetrate cells via a four-step mechanism. FEBS Lett 346: 92–98.

74. MurphyJR (2011) Mechanism of Diphtheria Toxin Catalytic Domain Delivery to the Eukaryotic Cell Cytosol and the Cellular Factors that Directly Participate in the Process. Toxins (Basel) 3: 294–308.

75. KnustZ, BlumenthalB, AktoriesK, SchmidtG (2009) Cleavage of Escherichia coli cytotoxic necrotizing factor 1 is required for full biologic activity. Infect Immun 77: 1835–1841.

76. RupnikM, PabstS, von Eichel-StreiberC, UrlaubH, SolingHD (2005) Characterization of the cleavage site and function of resulting cleavage fragments after limited proteolysis of Clostridium difficile toxin B (TcdB) by host cells. Microbiology 151: 199–208.

77. KrieglsteinKG, HenschenAH, WellerU, HabermannE (1991) Limited proteolysis of tetanus toxin. Relation to activity and identification of cleavage sites. Eur J Biochem 202: 41–51.

78. PirazziniM, RossettoO, BologneseP, ShoneCC, MontecuccoC (2011) Double anchorage to the membrane and intact inter-chain disulfide bond are required for the low pH induced entry of tetanus and botulinum neurotoxins into neurons. Cell Microbiol 13: 1731–1743.

79. FalnesPO, AriansenS, SandvigK, OlsnesS (2000) Requirement for prolonged action in the cytosol for optimal protein synthesis inhibition by diphtheria toxin. J Biol Chem 275: 4363–4368.

80. Morlon-GuyotJ, MereJ, BonhoureA, BeaumelleB (2009) Processing of Pseudomonas aeruginosa exotoxin A is dispensable for cell intoxication. Infect Immun 77: 3090–3099.

81. AlamiMr, TaupiacM-P, ReggioH, BienvenüeA, BeaumelleB (1998) Involvement of ATP-dependent Pseudomonas Exotoxin Translocation from a Late Recycling Compartment in Lymphocyte Intoxication Procedure. Mol Biol Cell 9: 387–402.

82. StavrinidesJ, MaW, GuttmanDS (2006) Terminal reassortment drives the quantum evolution of type III effectors in bacterial pathogens. PLoS Pathog 2: e104.

83. MiroldS, EhrbarK, WeissmullerA, PragerR, TschapeH, et al. (2001) Salmonella host cell invasion emerged by acquisition of a mosaic of separate genetic elements, including Salmonella pathogenicity island 1 (SPI1), SPI5, and sopE2. J Bacteriol 183: 2348–2358.

84. KrzywinskaE, KrzywinskiJ, SchoreyJS (2004) Naturally occurring horizontal gene transfer and homologous recombination in Mycobacterium. Microbiology 150: 1707–1712.

85. HaakeDA, SuchardMA, KelleyMM, DundooM, AltDP, et al. (2004) Molecular evolution and mosaicism of leptospiral outer membrane proteins involves horizontal DNA transfer. J Bacteriol 186: 2818–2828.

86. DaviesRL, CampbellS, WhittamTS (2002) Mosaic structure and molecular evolution of the leukotoxin operon (lktCABD) in Mannheimia (Pasteurella) haemolytica, Mannheimia glucosida, and Pasteurella trehalosi. J Bacteriol 184: 266–277.

87. BaldoL, LoN, WerrenJH (2005) Mosaic nature of the wolbachia surface protein. J Bacteriol 187: 5406–5418.

88. TonelloF, NalettoL, RomanelloV, Dal MolinF, MontecuccoC (2004) Tyrosine-728 and glutamic acid-735 are essential for the metalloproteolytic activity of the lethal factor of Bacillus anthracis. Biochem Biophys Res Commun 313: 496–502.

89. AfonsoA, LousadaS, SilvaJ, EllisAE, SilvaMT (1998) Neutrophil and macrophage responses to inflammation in the peritoneal cavity of rainbow trout Oncorhynchus mykiss. A light and electron microscopic cytochemical study. Dis Aquat Organ 34: 27–37.

90. do ValeA, AfonsoA, SilvaMT (2002) The professional phagocytes of sea bass (Dicentrarchus labrax L.): cytochemical characterisation of neutrophils and macrophages in the normal and inflamed peritoneal cavity. Fish Shellfish Immunol 13: 183–198.

91. BendtsenJD, NielsenH, von HeijneG, BrunakS (2004) Improved prediction of signal peptides: SignalP 3.0. J Mol Biol 340: 783–795.

92. NielsenH, EngelbrechtJ, BrunakS, von HeijneG (1997) Identification of prokaryotic and eukaryotic signal peptides and prediction of their cleavage sites. Protein Eng 10: 1–6.