Metalloprotease NleC Suppresses Host NF-κB/Inflammatory Responses by Cleaving p65 and Interfering with the p65/RPS3 Interaction

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The nuclear factor kappaB (NF-κB) signaling pathway is crucial for host defense, as it orchestrates both innate and adaptive immune responses. Beyond the best-studied Rel proteins (p65, RelB, c-Rel, p50 and p52), RPS3 has been recently identified as a “specifier” component of NF-κB, modulating the promoter selectivity and transcriptional specificity of NF-κB. In particular, the RPS3/p65-conferred signaling pathway was recently shown to play a critical role in host proinflammatory transcription and immune responses. Attaching and effacing (A/E) pathogens and others have acquired sophisticated mechanisms to modulate host NF-κB signaling pathways. We have found that NleC, a metalloprotease effector secreted by A/E pathogens, modulates host NF-κB signaling and inflammatory responses through a novel mechanism. NleC specifically recognizes and cleaves a small percentage of p65 and the generated N-terminal fragment of p65 interferes with the p65/RPS3 interaction, thereby amplifying the effect of cleaving only a small percentage of p65 molecules to selectively inhibit NF-κB gene expression. Our findings highlight a previously unappreciated mechanism through which pathogen-encoded proteases interfere with signaling cascades and inflammatory responses in host cells.

Vyšlo v časopise: Metalloprotease NleC Suppresses Host NF-κB/Inflammatory Responses by Cleaving p65 and Interfering with the p65/RPS3 Interaction. PLoS Pathog 11(3): e32767. doi:10.1371/journal.ppat.1004705
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1004705

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Zdroje

1. Clarke SC (2001) Diarrhoeagenic Escherichia coli—an emerging problem? Diagnostic microbiology and infectious disease 41 : 93–98. 11750160

2. Kaper JB, Nataro JP, Mobley HL (2004) Pathogenic Escherichia coli. Nature reviews Microbiology 2 : 123–140. 15040260

3. Donnenberg MS, Kaper JB, Finlay BB (1997) Interactions between enteropathogenic Escherichia coli and host epithelial cells. Trends in microbiology 5 : 109–114. 9080609

4. Hueck CJ (1998) Type III protein secretion systems in bacterial pathogens of animals and plants. Microbiology and molecular biology reviews: MMBR 62 : 379–433. 9618447

5. Coburn B, Sekirov I, Finlay BB (2007) Type III secretion systems and disease. Clinical microbiology reviews 20 : 535–549. 17934073

6. Deng W, Puente JL, Gruenheid S, Li Y, Vallance BA, et al. (2004) Dissecting virulence: systematic and functional analyses of a pathogenicity island. Proceedings of the National Academy of Sciences of the United States of America 101 : 3597–3602. 14988506

7. Tobe T, Beatson SA, Taniguchi H, Abe H, Bailey CM, et al. (2006) An extensive repertoire of type III secretion effectors in Escherichia coli O157 and the role of lambdoid phages in their dissemination. Proceedings of the National Academy of Sciences of the United States of America 103 : 14941–14946. 16990433

8. Garcia-Angulo VA, Deng W, Thomas NA, Finlay BB, Puente JL (2008) Regulation of expression and secretion of NleH, a new non-locus of enterocyte effacement-encoded effector in Citrobacter rodentium. Journal of bacteriology 190 : 2388–2399. doi: 10.1128/JB.01602-07 18223087

9. Deng W, Yu HB, de Hoog CL, Stoynov N, Li Y, et al. (2012) Quantitative Proteomic Analysis of Type III Secretome of Enteropathogenic Escherichia coli Reveals an Expanded Effector Repertoire for Attaching/Effacing Bacterial Pathogens. Molecular & cellular proteomics: MCP 11 : 692–709.

10. Deng W, de Hoog CL, Yu HB, Li Y, Croxen MA, et al. (2010) A comprehensive proteomic analysis of the type III secretome of Citrobacter rodentium. The Journal of biological chemistry 285 : 6790–6800. doi: 10.1074/jbc.M109.086603 20034934

11. Nadler C, Baruch K, Kobi S, Mills E, Haviv G, et al. (2010) The type III secretion effector NleE inhibits NF-kappaB activation. PLoS pathogens 6: e1000743. doi: 10.1371/journal.ppat.1000743 20126447

12. Royan SV, Jones RM, Koutsouris A, Roxas JL, Falzari K, et al. (2010) Enteropathogenic E. coli non-LEE encoded effectors NleH1 and NleH2 attenuate NF-kappaB activation. Molecular microbiology 78 : 1232–1245. doi: 10.1111/j.1365-2958.2010.07400.x 21091507

13. Newton HJ, Pearson JS, Badea L, Kelly M, Lucas M, et al. (2010) The type III effectors NleE and NleB from enteropathogenic E. coli and OspZ from Shigella block nuclear translocation of NF-kappaB p65. PLoS pathogens 6: e1000898. doi: 10.1371/journal.ppat.1000898 20485572

14. Baruch K, Gur-Arie L, Nadler C, Koby S, Yerushalmi G, et al. (2011) Metalloprotease type III effectors that specifically cleave JNK and NF-kappaB. Embo J 30 : 221–231. doi: 10.1038/emboj.2010.297 21113130

15. Vossenkamper A, Marches O, Fairclough PD, Warnes G, Stagg AJ, et al. (2010) Inhibition of NF-kappaB signaling in human dendritic cells by the enteropathogenic Escherichia coli effector protein NleE. Journal of immunology 185 : 4118–4127. doi: 10.4049/jimmunol.1000500 20833837

16. 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 pathogens 6: e1001231. doi: 10.1371/journal.ppat.1001231 21187904

17. Gao X, Wan F, Mateo K, Callegari E, Wang D, et al. (2009) Bacterial effector binding to ribosomal protein s3 subverts NF-kappaB function. PLoS pathogens 5: e1000708. doi: 10.1371/journal.ppat.1000708 20041225

18. Wan F, Weaver A, Gao X, Bern M, Hardwidge PR, et al. (2011) IKKbeta phosphorylation regulates RPS3 nuclear translocation and NF-kappaB function during infection with Escherichia coli strain O157:H7. Nature immunology 12 : 335–343. doi: 10.1038/ni.2007 21399639

19. Pham TH, Gao X, Tsai K, Olsen R, Wan F, et al. (2012) Functional differences and interactions between the Escherichia coli type III secretion system effectors NleH1 and NleH2. Infection and immunity 80 : 2133–2140. doi: 10.1128/IAI.06358-11 22451523

20. Sham HP, Shames SR, Croxen MA, Ma C, Chan JM, et al. (2011) Attaching and effacing bacterial effector NleC suppresses epithelial inflammatory responses by inhibiting NF-kappaB and p38 mitogen-activated protein kinase activation. Infection and immunity 79 : 3552–3562. doi: 10.1128/IAI.05033-11 21746856

21. Le Negrate G (2012) Subversion of innate immune responses by bacterial hindrance of NF-kappaB pathway. Cellular microbiology 14 : 155–167. doi: 10.1111/j.1462-5822.2011.01719.x 22044780

22. Santoro MG, Rossi A, Amici C (2003) NF-kappaB and virus infection: who controls whom. Embo J 22 : 2552–2560. 12773372

23. Mulhern O, Harrington B, Bowie AG (2009) Modulation of innate immune signalling pathways by viral proteins. Advances in experimental medicine and biology 666 : 49–63. 20054974

24. Rahman MM, McFadden G (2006) Modulation of tumor necrosis factor by microbial pathogens. PLoS pathogens 2: e4. 16518473

25. Rahman MM, McFadden G (2011) Modulation of NF-kappaB signalling by microbial pathogens. Nature reviews Microbiology 9 : 291–306. doi: 10.1038/nrmicro2539 21383764

26. O’Callaghan D, Stebbins CE (2010) Host-microbe interactions: bacteria. Current opinion in microbiology 13 : 1–3. doi: 10.1016/j.mib.2009.12.006 20044300

27. Lilic M, Galkin VE, Orlova A, VanLoock MS, Egelman EH, et al. (2003) Salmonella SipA polymerizes actin by stapling filaments with nonglobular protein arms. Science 301 : 1918–1921. 14512630

28. Clements A, Young J, Constantinou N, Frankel G (2012) Infection strategies of enteric pathogenic E. coli. Gut microbes 3.

29. Muhlen S, Ruchaud-Sparagano MH, Kenny B (2011) Proteasome-independent degradation of canonical NFkappaB complex components by the NleC protein of pathogenic Escherichia coli. The Journal of biological chemistry 286 : 5100–5107. doi: 10.1074/jbc.M110.172254 21148319

30. Pearson JS, Riedmaier P, Marches O, Frankel G, Hartland EL (2011) A type III effector protease NleC from enteropathogenic Escherichia coli targets NF-kappaB for degradation. Molecular microbiology 80 : 219–230. doi: 10.1111/j.1365-2958.2011.07568.x 21306441

31. Ruchaud-Sparagano MH, Maresca M, Kenny B (2007) Enteropathogenic Escherichia coli (EPEC) inactivate innate immune responses prior to compromising epithelial barrier function. Cellular microbiology 9 : 1909–1921. 17388785

32. Hayden MS, Ghosh S (2004) Signaling to NF-kappaB. Genes & development 18 : 2195–2224.

33. Wan F, Lenardo MJ (2009) Specification of DNA binding activity of NF-kappaB proteins. Cold Spring Harbor perspectives in biology 1: a000067. doi: 10.1101/cshperspect.a000067 20066093

34. Wan F, Anderson DE, Barnitz RA, Snow A, Bidere N, et al. (2007) Ribosomal protein S3: a KH domain subunit in NF-kappaB complexes that mediates selective gene regulation. Cell 131 : 927–939. 18045535

35. Fu K, Sun X, Zheng W, Wier EM, Hodgson A, et al. (2013) Sam68 modulates the promoter specificity of NF-kappaB and mediates expression of CD25 in activated T cells. Nature communications 4 : 1909. doi: 10.1038/ncomms2916 23715268

36. Alkalay I, Yaron A, Hatzubai A, Orian A, Ciechanover A, et al. (1995) Stimulation-dependent I kappa B alpha phosphorylation marks the NF-kappa B inhibitor for degradation via the ubiquitin-proteasome pathway. Proceedings of the National Academy of Sciences of the United States of America 92 : 10599–10603. 7479848

37. Cadera EJ, Wan F, Amin RH, Nolla H, Lenardo MJ, et al. (2009) NF-kappaB activity marks cells engaged in receptor editing. The Journal of experimental medicine 206 : 1803–1816. doi: 10.1084/jem.20082815 19581408

38. Mokhtari D, Barbu A, Mehmeti I, Vercamer C, Welsh N (2009) Overexpression of the nuclear factor-kappaB subunit c-Rel protects against human islet cell death in vitro. American journal of physiology Endocrinology and metabolism 297: E1067–1077. doi: 10.1152/ajpendo.00212.2009 19706790

39. Sen N, Paul BD, Gadalla MM, Mustafa AK, Sen T, et al. (2012) Hydrogen sulfide-linked sulfhydration of NF-kappaB mediates its antiapoptotic actions. Mol Cell 45 : 13–24. doi: 10.1016/j.molcel.2011.10.021 22244329

40. Wier EM, Neighoff J, Sun X, Fu K, Wan F (2012) Identification of an N-terminal truncation of the NF-kappaB p65 subunit that specifically modulates ribosomal protein S3-dependent NF-kappaB gene expression. The Journal of biological chemistry 287 : 43019–43029. doi: 10.1074/jbc.M112.388694 23115242

41. Baruch K, Gur-Arie L, Nadler C, Koby S, Yerushalmi G, et al. (2011) Metalloprotease type III effectors that specifically cleave JNK and NF-kappaB. The EMBO journal 30 : 221–231. doi: 10.1038/emboj.2010.297 21113130

42. Shames SR, Bhavsar AP, Croxen MA, Law RJ, Mak SH, et al. (2011) The pathogenic Escherichia coli type III secreted protease NleC degrades the host acetyltransferase p300. Cellular microbiology 13 : 1542–1557. doi: 10.1111/j.1462-5822.2011.01640.x 21812888

43. Vallabhapurapu S, Karin M (2009) Regulation and function of NF-kappaB transcription factors in the immune system. Annu Rev Immunol 27 : 693–733. doi: 10.1146/annurev.immunol.021908.132641 19302050

44. Li W, Liu Y, Sheng X, Yin P, Hu F, et al. (2014) Structure and mechanism of a type III secretion protease, NleC. Acta crystallographica Section D, Biological crystallography 70 : 40–47. doi: 10.1107/S1399004713024619 24419377

45. Turco MM, Sousa MC (2014) The structure and specificity of the type III secretion system effector NleC suggest a DNA mimicry mechanism of substrate recognition. Biochemistry 53 : 5131–5139. doi: 10.1021/bi500593e 25040221

46. Sorimachi H, Mamitsuka H, Ono Y (2012) Understanding the substrate specificity of conventional calpains. Biological chemistry 393 : 853–871. doi: 10.1515/hsz-2012-0143 22944687

47. Chen FE, Huang DB, Chen YQ, Ghosh G (1998) Crystal structure of p50/p65 heterodimer of transcription factor NF-kappaB bound to DNA. Nature 391 : 410–413. 9450761

48. Mundy R, MacDonald TT, Dougan G, Frankel G, Wiles S (2005) Citrobacter rodentium of mice and man. Cellular microbiology 7 : 1697–1706. 16309456

49. Wan F, Lenardo MJ (2010) The nuclear signaling of NF-kappaB: current knowledge, new insights, and future perspectives. Cell research 20 : 24–33. doi: 10.1038/cr.2009.137 19997086

50. Shames SR, Finlay BB (2012) Bacterial effector interplay: a new way to view effector function. Trends in microbiology 20 : 214–219. doi: 10.1016/j.tim.2012.02.007 22425230

51. Wong AR, Pearson JS, Bright MD, Munera D, Robinson KS, et al. (2011) Enteropathogenic and enterohaemorrhagic Escherichia coli: even more subversive elements. Molecular microbiology 80 : 1420–1438. doi: 10.1111/j.1365-2958.2011.07661.x 21488979

52. Silva DS, Pereira LM, Moreira AR, Ferreira-da-Silva F, Brito RM, et al. (2013) The apoptogenic toxin AIP56 is a metalloprotease A-B toxin that cleaves NF-kappab P65. PLoS pathogens 9: e1003128. doi: 10.1371/journal.ppat.1003128 23468618

53. Doyle PS, Zhou YM, Hsieh I, Greenbaum DC, McKerrow JH, et al. (2011) The Trypanosoma cruzi protease cruzain mediates immune evasion. PLoS pathogens 7: e1002139. doi: 10.1371/journal.ppat.1002139 21909255

54. 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. The Journal of biological chemistry 285 : 41320–41327. doi: 10.1074/jbc.M110.152280 21041296

55. 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. The Journal of biological chemistry 280 : 24153–24158. 15845545

56. Cameron P, McGachy A, Anderson M, Paul A, Coombs GH, et al. (2004) Inhibition of lipopolysaccharide-induced macrophage IL-12 production by Leishmania mexicana amastigotes: the role of cysteine peptidases and the NF-kappaB signaling pathway. Journal of immunology 173 : 3297–3304. 15322192

57. Coiras M, Lopez-Huertas MR, Mateos E, Alcami J (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. doi: 10.1186/1742-4690-5-109 19046417

58. Lad SP, Yang G, Scott DA, Wang G, Nair P, et al. (2007) Chlamydial CT441 is a PDZ domain-containing tail-specific protease that interferes with the NF-kappaB pathway of immune response. Journal of bacteriology 189 : 6619–6625. 17631635

59. Lad SP, Li J, da Silva Correia J, Pan Q, Gadwal S, et al. (2007) Cleavage of p65/RelA of the NF-kappaB pathway by Chlamydia. Proceedings of the National Academy of Sciences of the United States of America 104 : 2933–2938. 17301240

60. Dooher JE, Paz-Priel I, Houng S, Baldwin AS Jr, Friedman AD (2011) C/EBPalpha, C/EBPalpha oncoproteins, or C/EBPbeta preferentially bind NF-kappaB p50 compared with p65, focusing therapeutic targeting on the C/EBP:p50 interaction. Molecular cancer research: MCR 9 : 1395–1405. doi: 10.1158/1541-7786.MCR-11-0072 21813505

61. Flint N, Cove FL, Evans GS (1991) A low-temperature method for the isolation of small-intestinal epithelium along the crypt-villus axis. The Biochemical journal 280 (Pt 2): 331–334. 1747105

62. Childs WC 3rd, Gibbons RJ (1988) Use of Percoll density gradients for studying the attachment of bacteria to oral epithelial cells. Journal of dental research 67 : 826–830. 3163351

63. Colombo AV, Silva CM, Haffajee A, Colombo AP (2006) Identification of oral bacteria associated with crevicular epithelial cells from chronic periodontitis lesions. Journal of medical microbiology 55 : 609–615. 16585650

64. Izhar M, Nuchamowitz Y, Mirelman D (1982) Adherence of Shigella flexneri to guinea pig intestinal cells is mediated by a mucosal adhesion. Infection and immunity 35 : 1110–1118. 7040246

65. Qualls JE, Kaplan AM, van Rooijen N, Cohen DA (2006) Suppression of experimental colitis by intestinal mononuclear phagocytes. Journal of leukocyte biology 80 : 802–815. 16888083