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Hyperinvasive Meningococci Induce Intra-nuclear Cleavage of the NF-κB Protein p65/RelA by Meningococcal IgA Protease


Strains of Neisseria meningitidis isolated from patients induce apoptotic cell death, whereas strains isolated from healthy carriage isolates do not. Part of the difference has been shown to arise from differential modulation of NF-κB during meningococcal infection. While non-invasive isolates of Neisseria meningitidis provoke a sustained NF-κB activation in epithelial cells, hyperinvasive isolates only induce an early NF-κB activation followed by a sustained activation of JNK and apoptosis. Here, we elucidate the mechanism conferring this differential modulation, specifically showing that ST-11 hyperinvasive isolates promote specific cleavage of NF-κB p65/RelA component in a manner dependent on the secreted IgA protease. This cleavage occurs within the nuclear compartment. Secreted IgA protease from non-invasive isolates was unable to reach the nuclear compartment of infected cells, resulting in a sustained activation of NF-κB activity and subsequent cytoprotective effect. Modulation of NF-κB-related signaling is likely a double-edged sword to decide the fate of meningococcal infection.


Vyšlo v časopise: Hyperinvasive Meningococci Induce Intra-nuclear Cleavage of the NF-κB Protein p65/RelA by Meningococcal IgA Protease. PLoS Pathog 11(8): e32767. doi:10.1371/journal.ppat.1005078
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1005078

Souhrn

Strains of Neisseria meningitidis isolated from patients induce apoptotic cell death, whereas strains isolated from healthy carriage isolates do not. Part of the difference has been shown to arise from differential modulation of NF-κB during meningococcal infection. While non-invasive isolates of Neisseria meningitidis provoke a sustained NF-κB activation in epithelial cells, hyperinvasive isolates only induce an early NF-κB activation followed by a sustained activation of JNK and apoptosis. Here, we elucidate the mechanism conferring this differential modulation, specifically showing that ST-11 hyperinvasive isolates promote specific cleavage of NF-κB p65/RelA component in a manner dependent on the secreted IgA protease. This cleavage occurs within the nuclear compartment. Secreted IgA protease from non-invasive isolates was unable to reach the nuclear compartment of infected cells, resulting in a sustained activation of NF-κB activity and subsequent cytoprotective effect. Modulation of NF-κB-related signaling is likely a double-edged sword to decide the fate of meningococcal infection.


Zdroje

1. Tzeng YL, Stephens DS (2000) Epidemiology and pathogenesis of Neisseria meningitidis. Microbes Infect 2: 687–700. 10884620

2. Yazdankhah SP, Caugant DA (2004) Neisseria meningitidis: an overview of the carriage state. J Med Microbiol 53: 821–832. 15314188

3. Densen P (1989) Interaction of complement with Neisseria meningitidis and Neisseria gonorrhoeae. Clin Microbiol Rev 2 Suppl: S11–17. 2497954

4. Emonts M, Hazelzet JA, de Groot R, Hermans PW (2003) Host genetic determinants of Neisseria meningitidis infections. Lancet Infect Dis 3: 565–577. 12954563

5. Goldschneider I, Gotschlich EC, Artenstein MS (1969) Human immunity to the meningococcus. II. Development of natural immunity. J Exp Med 129: 1327–1348. 4977281

6. Caugant DA, Kristiansen BE, Froholm LO, Bovre K, Selander RK (1988) Clonal diversity of Neisseria meningitidis from a population of asymptomatic carriers. Infect Immun 56: 2060–2068. 3135270

7. Jolley KA, Kalmusova J, Feil EJ, Gupta S, Musilek M, et al. (2000) Carried meningococci in the Czech Republic: a diverse recombining population. J Clin Microbiol 38: 4492–4498. 11101585

8. Watkins ER, Maiden MC (2012) Persistence of hyperinvasive meningococcal strain types during global spread as recorded in the PubMLST database. PLoS ONE 7: e45349. doi: 10.1371/journal.pone.0045349 23028953

9. Deghmane AE, Parent du Chatelet I, Szatanik M, Hong E, Ruckly C, et al. (2010) Emergence of new virulent Neisseria meningitidis serogroup C sequence type 11 isolates in France. J Infect Dis 202: 247–250. doi: 10.1086/653583 20515410

10. Perrocheau A, Taha M, Levy-Bruhl D (2005) Epidemiology of invasive meningococcal disease in France in 2003. Euro Surveill 10.

11. Levy-Bruhl D, Perrocheau A, Mora M, Taha MK, Dromell-Chabrier S, et al. (2002) Vaccination campaign following an increase in incidence of serogroup C meningococcal diseases in the department of Puy-de-Dome (France). Euro Surveill 7: 74–76. 12631934

12. Stephens DS, Farley MM (1991) Pathogenic events during infection of the human nasopharynx with Neisseria meningitidis and Haemophilus influenzae. Rev Infect Dis 13: 22–33. 1901998

13. Birkness KA, Swisher BL, White EH, Long EG, Ewing EP Jr., et al. (1995) A tissue culture bilayer model to study the passage of Neisseria meningitidis. Infect Immun 63: 402–409. 7822003

14. Klein NJ, Ison CA, Peakman M, Levin M, Hammerschmidt S, et al. (1996) The influence of capsulation and lipooligosaccharide structure on neutrophil adhesion molecule expression and endothelial injury by Neisseria meningitidis. J Infect Dis 173: 172–179. 8537655

15. Warren HS Jr., Gonzalez RG, Tian D (2003) Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 38–2003. A 12-year-old girl with fever and coma. N Engl J Med 349: 2341–2349. 14668461

16. Zarantonelli ML, Lancellotti M, Deghmane AE, Giorgini D, Hong E, et al. (2008) Hyperinvasive genotypes of Neisseria meningitidis in France. Clin Microbiol Infect 14: 467–472. doi: 10.1111/j.1469-0691.2008.01955.x 18294240

17. Deghmane AE, Veckerle C, Giorgini D, Hong E, Ruckly C, et al. (2009) Differential modulation of TNF-alpha-induced apoptosis by Neisseria meningitidis. PLoS Pathog 5: e1000405. doi: 10.1371/journal.ppat.1000405 19412525

18. Deghmane AE, El Kafsi H, Giorgini D, Abaza A, Taha MK (2011) Late repression of NF-kappaB activity by invasive but not non-invasive meningococcal isolates is required to display apoptosis of epithelial cells. PLoS Pathog 7: e1002403. doi: 10.1371/journal.ppat.1002403 22144896

19. Rothwarf DM, Karin M (1999) The NF-kappa B activation pathway: a paradigm in information transfer from membrane to nucleus. Sci STKE 1999: RE1. 11865184

20. Hoffmann A, Levchenko A, Scott ML, Baltimore D (2002) The IkappaB-NF-kappaB signaling module: temporal control and selective gene activation. Science 298: 1241–1245. 12424381

21. Mogensen TH, Paludan SR (2001) Molecular pathways in virus-induced cytokine production. Microbiol Mol Biol Rev 65: 131–150. 11238989

22. Rawlings ND, Waller M, Barrett AJ, Bateman A (2014) MEROPS: the database of proteolytic enzymes, their substrates and inhibitors. Nucleic Acids Res 42: D503–509. doi: 10.1093/nar/gkt953 24157837

23. Plaut AG, Bachovchin WW (1994) IgA-specific prolyl endopeptidases: serine type. Methods Enzymol 244: 137–151. 7845203

24. Hauck CR, Meyer TF (1997) The lysosomal/phagosomal membrane protein h-lamp-1 is a target of the IgA1 protease of Neisseria gonorrhoeae. FEBS Lett 405: 86–90. 9094430

25. Binscheck T, Bartels F, Bergel H, Bigalke H, Yamasaki S, et al. (1995) IgA protease from Neisseria gonorrhoeae inhibits exocytosis in bovine chromaffin cells like tetanus toxin. J Biol Chem 270: 1770–1774. 7829513

26. Grijpstra J, Arenas J, Rutten L, Tommassen J (2013) Autotransporter secretion: varying on a theme. Res Microbiol 164: 562–582. doi: 10.1016/j.resmic.2013.03.010 23567321

27. van Ulsen P, Rahman S, Jong WS, Daleke-Schermerhorn MH, Luirink J (2014) Type V secretion: from biogenesis to biotechnology. Biochim Biophys Acta 1843: 1592–1611. doi: 10.1016/j.bbamcr.2013.11.006 24269841

28. Pohlner J, Halter R, Beyreuther K, Meyer TF (1987) Gene structure and extracellular secretion of Neisseria gonorrhoeae IgA protease. Nature 325: 458–462. 3027577

29. Pohlner J, Langenberg U, Wolk U, Beck SC, Meyer TF (1995) Uptake and nuclear transport of Neisseria IgA1 protease-associated alpha-proteins in human cells. Mol Microbiol 17: 1073–1083. 8594327

30. van Ulsen P, van Alphen L, ten Hove J, Fransen F, van der Ley P, et al. (2003) A Neisserial autotransporter NalP modulating the processing of other autotransporters. Mol Microbiol 50: 1017–1030. 14617158

31. Irmler M, Thome M, Hahne M, Schneider P, Hofmann K, et al. (1997) Inhibition of death receptor signals by cellular FLIP. Nature 388: 190–195. 9217161

32. Weinrauch Y, Zychlinsky A (1999) The induction of apoptosis by bacterial pathogens. Annu Rev Microbiol 53: 155–187. 10547689

33. Hersh D, Monack DM, Smith MR, Ghori N, Falkow S, et al. (1999) The Salmonella invasin SipB induces macrophage apoptosis by binding to caspase-1. Proc Natl Acad Sci U S A 96: 2396–2401. 10051653

34. Royer PJ, Rogers AJ, Wooldridge KG, Tighe P, Mahdavi J, et al. (2013) Deciphering the contribution of human meningothelial cells to the inflammatory and antimicrobial response at the meninges. Infect Immun 81: 4299–4310. doi: 10.1128/IAI.00477-13 24002066

35. Zhang SQ, Kovalenko A, Cantarella G, Wallach D (2000) Recruitment of the IKK signalosome to the p55 TNF receptor: RIP and A20 bind to NEMO (IKKgamma) upon receptor stimulation. Immunity 12: 301–311. 10755617

36. Mauro C, Pacifico F, Lavorgna A, Mellone S, Iannetti A, et al. (2006) ABIN-1 binds to NEMO/IKKgamma and co-operates with A20 in inhibiting NF-kappaB. J Biol Chem 281: 18482–18488. 16684768

37. Skaug B, Chen J, Du F, He J, Ma A, et al. (2011) Direct, noncatalytic mechanism of IKK inhibition by A20. Mol Cell 44: 559–571. doi: 10.1016/j.molcel.2011.09.015 22099304

38. Wertz IE, O'Rourke KM, Zhou H, Eby M, Aravind L, et al. (2004) De-ubiquitination and ubiquitin ligase domains of A20 downregulate NF-kappaB signalling. Nature 430: 694–699. 15258597

39. Wang Y, Xiang GS, Kourouma F, Umar S (2006) Citrobacter rodentium-induced NF-kappaB activation in hyperproliferating colonic epithelia: role of p65 (Ser536) phosphorylation. Br J Pharmacol 148: 814–824. 16751795

40. Chen Y, Jin H, Chen P, Li Z, Meng X, et al. (2012) Haemophilus parasuis infection activates the NF-kappaB pathway in PK-15 cells through IkappaB degradation. Vet Microbiol 160: 259–263. doi: 10.1016/j.vetmic.2012.05.021 22704560

41. Kumar A, Zhang J, Yu FS (2004) Innate immune response of corneal epithelial cells to Staphylococcus aureus infection: role of peptidoglycan in stimulating proinflammatory cytokine secretion. Invest Ophthalmol Vis Sci 45: 3513–3522. 15452057

42. Wei L, Kwang J, Wang J, Shi L, Yang B, et al. (2008) Porcine circovirus type 2 induces the activation of nuclear factor kappa B by IkappaBalpha degradation. Virology 378: 177–184. doi: 10.1016/j.virol.2008.05.013 18561971

43. Baldwin AS Jr. (1996) The NF-kappa B and I kappa B proteins: new discoveries and insights. Annu Rev Immunol 14: 649–683. 8717528

44. Gilmore TD, Koedood M, Piffat KA, White DW (1996) Rel/NF-kappaB/IkappaB proteins and cancer. Oncogene 13: 1367–1378. 8875974

45. O'Neill LA, Kaltschmidt C (1997) NF-kappa B: a crucial transcription factor for glial and neuronal cell function. Trends Neurosci 20: 252–258. 9185306

46. Schmidt-Ullrich R, Aebischer T, Hulsken J, Birchmeier W, Klemm U, et al. (2001) Requirement of NF-kappaB/Rel for the development of hair follicles and other epidermal appendices. Development 128: 3843–3853. 11585809

47. Zawia NH, Sharan R, Brydie M, Oyama T, Crumpton T (1998) Sp1 as a target site for metal-induced perturbations of transcriptional regulation of developmental brain gene expression. Brain Res Dev Brain Res 107: 291–298. 9593950

48. Arenas J, Nijland R, Rodriguez FJ, Bosma TN, Tommassen J (2013) Involvement of three meningococcal surface-exposed proteins, the heparin-binding protein NhbA, the alpha-peptide of IgA protease and the autotransporter protease NalP, in initiation of biofilm formation. Mol Microbiol 87: 254–268. doi: 10.1111/mmi.12097 23163582

49. Krause A, Holtmann H, Eickemeier S, Winzen R, Szamel M, et al. (1998) Stress-activated protein kinase/Jun N-terminal kinase is required for interleukin (IL)-1-induced IL-6 and IL-8 gene expression in the human epidermal carcinoma cell line KB. J Biol Chem 273: 23681–23689. 9726973

50. Ip YT, Davis RJ (1998) Signal transduction by the c-Jun N-terminal kinase (JNK)—from inflammation to development. Curr Opin Cell Biol 10: 205–219. 9561845

51. Chu WM, Ostertag D, Li ZW, Chang L, Chen Y, et al. (1999) JNK2 and IKKbeta are required for activating the innate response to viral infection. Immunity 11: 721–731. 10626894

52. Kamata H, Honda S, Maeda S, Chang L, Hirata H, et al. (2005) Reactive oxygen species promote TNFalpha-induced death and sustained JNK activation by inhibiting MAP kinase phosphatases. Cell 120: 649–661. 15766528

53. Bhavsar AP, D'Elia MA, Sahakian TD, Brown ED (2007) The Amino terminus of Bacillus subtilis TagB possesses separable localization and functional properties. J Bacteriol 189: 6816–6823. 17660278

54. Moon DC, Gurung M, Lee JH, Lee YS, Choi CW, et al. (2012) Screening of nuclear targeting proteins in Acinetobacter baumannii based on nuclear localization signals. Res Microbiol 163: 279–285. doi: 10.1016/j.resmic.2012.02.001 22366694

55. McSweeney LA, Dreyfus LA (2004) Nuclear localization of the Escherichia coli cytolethal distending toxin CdtB subunit. Cell Microbiol 6: 447–458. 15056215

56. Lara-Tejero M, Galan JE (2000) A bacterial toxin that controls cell cycle progression as a deoxyribonuclease I-like protein. Science 290: 354–357. 11030657

57. Choi CH, Hyun SH, Kim J, Lee YC, Seol SY, et al. (2008) Nuclear translocation and DNAse I-like enzymatic activity of Acinetobacter baumannii outer membrane protein A. FEMS Microbiol Lett 288: 62–67. doi: 10.1111/j.1574-6968.2008.01323.x 18783439

58. Moon DC, Choi CH, Lee SM, Lee JH, Kim SI, et al. (2012) Nuclear translocation of Acinetobacter baumannii transposase induces DNA methylation of CpG regions in the promoters of E-cadherin gene. PLoS ONE 7: e38974. doi: 10.1371/journal.pone.0038974 22685614

59. Arbibe L, Kim DW, Batsche E, Pedron T, Mateescu B, et al. (2007) An injected bacterial effector targets chromatin access for transcription factor NF-kappaB to alter transcription of host genes involved in immune responses. Nat Immunol 8: 47–56. 17159983

60. Khairalla AS, Omer SA, Mahdavi J, Aslam A, Dufailu OA, et al. (2015) Nuclear trafficking, histone cleavage and induction of apoptosis by the meningococcal App and MspA autotransporters. Cell Microbiol.

61. Plaut AG, Gilbert JV, Artenstein MS, Capra JD (1975) Neisseria gonorrhoeae and neisseria meningitidis: extracellular enzyme cleaves human immunoglobulin A. Science 190: 1103–1105. 810892

62. Lin L, Ayala P, Larson J, Mulks M, Fukuda M, et al. (1997) The Neisseria type 2 IgA1 protease cleaves LAMP1 and promotes survival of bacteria within epithelial cells. Mol Microbiol 24: 1083–1094. 9220014

63. Ayala P, Vasquez B, Wetzler L, So M (2002) Neisseria gonorrhoeae porin P1.B induces endosome exocytosis and a redistribution of Lamp1 to the plasma membrane. Infect Immun 70: 5965–5971. 12379671

64. Senior BW, Stewart WW, Galloway C, Kerr MA (2001) Cleavage of the hormone human chorionic gonadotropin, by the Type 1 IgA1 protease of Neisseria gonorrhoeae, and its implications. J Infect Dis 184: 922–925. 11550129

65. Neznanov N, Chumakov KM, Neznanova L, Almasan A, Banerjee AK, et al. (2005) Proteolytic cleavage of the p65-RelA subunit of NF-kappaB during poliovirus infection. J Biol Chem 280: 24153–24158. 15845545

66. Yen H, Ooka T, Iguchi A, Hayashi T, Sugimoto N, et al. (2010) NleC, a type III secretion protease, compromises NF-kappaB activation by targeting p65/RelA. PLoS Pathog 6: e1001231. doi: 10.1371/journal.ppat.1001231 21187904

67. Christian J, Vier J, Paschen SA, Hacker G (2010) Cleavage of the NF-kappaB family protein p65/RelA by the chlamydial protease-like activity factor (CPAF) impairs proinflammatory signaling in cells infected with Chlamydiae. J Biol Chem 285: 41320–41327. doi: 10.1074/jbc.M110.152280 21041296

68. Dietrich G, Kurz S, Hubner C, Aepinus C, Theiss S, et al. (2003) Transcriptome analysis of Neisseria meningitidis during infection. J Bacteriol 185: 155–164. 12486052

69. Muller A, Gunther D, Dux F, Naumann M, Meyer TF, et al. (1999) Neisserial porin (PorB) causes rapid calcium influx in target cells and induces apoptosis by the activation of cysteine proteases. Embo J 18: 339–352. 9889191

70. Sjolinder M, Altenbacher G, Hagner M, Sun W, Schedin-Weiss S, et al. (2012) Meningococcal outer membrane protein NhhA triggers apoptosis in macrophages. PLoS ONE 7: e29586. doi: 10.1371/journal.pone.0029586 22238624

71. Halter R, Pohlner J, Meyer TF (1989) Mosaic-like organization of IgA protease genes in Neisseria gonorrhoeae generated by horizontal genetic exchange in vivo. Embo J 8: 2737–2744. 2511009

72. Lomholt H, Poulsen K, Kilian M (1995) Comparative characterization of the iga gene encoding IgA1 protease in Neisseria meningitidis, Neisseria gonorrhoeae and Haemophilus influenzae. Mol Microbiol 15: 495–506. 7783620

73. Jose J, Wolk U, Lorenzen D, Wenschuh H, Meyer TF (2000) Human T-cell response to meningococcal immunoglobulin A1 protease associated alpha-proteins. Scand J Immunol 51: 176–185. 10722372

74. Roussel-Jazede V, Arenas J, Langereis JD, Tommassen J, van Ulsen P (2014) Variable processing of the IgA protease autotransporter at the cell surface of Neisseria meningitidis. Microbiology 160: 2421–2431. doi: 10.1099/mic.0.082511-0 25161279

75. Stephens DS, Zimmer SM (2002) Pathogenesis, Therapy, and Prevention of Meningococcal Sepsis. Curr Infect Dis Rep 4: 377–386. 12228024

76. Raghunathan PL, Jones JD, Tiendrebeogo SR, Sanou I, Sangare L, et al. (2006) Predictors of immunity after a major serogroup W-135 meningococcal disease epidemic, Burkina Faso, 2002. J Infect Dis 193: 607–616. 16453255

77. Hanahan D (1983) Studies on transformation of Escherichia coli with plasmids. J Mol Biol 166: 557–580. 6345791

78. Studier FW, Rosenberg AH, Dunn JJ, Dubendorff JW (1990) Use of T7 RNA polymerase to direct expression of cloned genes. Methods Enzymol 185: 60–89. 2199796

79. Kellogg DS Jr., Peacock WL Jr., Deacon WE, Brown L, Pirkle DI (1963) Neisseria Gonorrhoeae. I. Virulence Genetically Linked to Clonal Variation. J Bacteriol 85: 1274–1279. 14047217

80. Jolley KA, Brehony C, Maiden MC (2007) Molecular typing of meningococci: recommendations for target choice and nomenclature. FEMS Microbiol Rev 31: 89–96. 17168996

81. Robinson K, Taraktsoglou M, Rowe KS, Wooldridge KG, Ala'Aldeen DA (2004) Secreted proteins from Neisseria meningitidis mediate differential human gene expression and immune activation. Cell Microbiol 6: 927–938. 15339268

82. Prentki P, Krisch HM (1984) In vitro insertional mutagenesis with a selectable DNA fragment. Gene 29: 303–313. 6237955

83. Nassif X, Puaoi D, So M (1991) Transposition of Tn1545-delta 3 in the pathogenic Neisseriae: a genetic tool for mutagenesis. J Bacteriol 173: 2147–2154. 1848839

84. Landt O, Grunert HP, Hahn U (1990) A general method for rapid site-directed mutagenesis using the polymerase chain reaction. Gene 96: 125–128. 2265750

85. Derre I, Rapoport G, Msadek T (1999) CtsR, a novel regulator of stress and heat shock response, controls clp and molecular chaperone gene expression in gram-positive bacteria. Mol Microbiol 31: 117–131. 9987115

86. Schmidt-Ullrich R, Memet S, Lilienbaum A, Feuillard J, Raphael M, et al. (1996) NF-kappaB activity in transgenic mice: developmental regulation and tissue specificity. Development 122: 2117–2128. 8681793

87. Deghmane AE, Giorgini D, Larribe M, Alonso JM, Taha MK (2002) Down-regulation of pili and capsule of Neisseria meningitidis upon contact with epithelial cells is mediated by CrgA regulatory protein. Mol Microbiol 43: 1555–1564. 11952904

88. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 25: 402–408. 11846609

89. Matin RN, Chikh A, Chong SL, Mesher D, Graf M, et al. (2013) p63 is an alternative p53 repressor in melanoma that confers chemoresistance and a poor prognosis. J Exp Med 210: 581–603. doi: 10.1084/jem.20121439 23420876

90. Deghmane AE, Soualhine H, Bach H, Sendide K, Itoh S, et al. (2007) Lipoamide dehydrogenase mediates retention of coronin-1 on BCG vacuoles, leading to arrest in phagosome maturation. J Cell Sci 120: 2796–2806. 17652161

91. Philpott DJ, Belaid D, Troubadour P, Thiberge JM, Tankovic J, et al. (2002) Reduced activation of inflammatory responses in host cells by mouse-adapted Helicobacter pylory isolates. Cell Microbiol 4: 285–296. 12064285

92. Vitovski S, Read RC, Sayers JR (1999) Invasive isolates of Neisseria meningitidis possess enhanced immunoglobulin A1 protease activity compared to colonizing strains. Faseb J 13: 331–337. 9973321

93. Hill M, Deghmane AE, Segovia M, Zarantonelli ML, Tilly G, et al. (2011) Penicillin binding proteins as danger signals: meningococcal penicillin binding protein 2 activates dendritic cells through Toll-like receptor 4. PLoS ONE 6: e23995. doi: 10.1371/journal.pone.0023995 22046231

94. Ducey TF, Carson MB, Orvis J, Stintzi AP, Dyer DW (2005) Identification of the iron-responsive genes of Neisseria gonorrhoeae by microarray analysis in defined medium. J Bacteriol 187: 4865–4874. 15995201

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