The Serine Protease EspC from Enteropathogenic Regulates Pore Formation and Cytotoxicity Mediated by the Type III Secretion System

English version České info

Enteropathogenic Escherichia coli (EPEC) is an important diarrheal pathogen responsible for infant diarrhoea associated with significant morbidity and mortality rates in developing countries. Upon ingestion EPEC colonizes the intestinal mucosa, causing characteristic lesions on enterocytes. Using a type III secretion system (T3SS) acting as a molecular syringe, EPEC injects numerous bacterial proteins into host cells that disrupt the intestinal epithelium homeostasis. Injection of T3SS proteins requires the insertion into the host cell plasma membrane of bacterial protein complex, called the "translocon", associated with pore-forming activity. In addition to the T3SS, EPEC also secretes other bacterial toxins involved in virulence. Among these, the EspC is a protease reported to degrade various host proteins. In this paper, we have characterized an "unsuspected role" for EspC. We show that EspC degrades the T3SS translocon components following cell contact and regulates T3SS-dependent pore formation in epithelial cells. The EspC control of pore formation limits cytotoxicity and thus, is expected to limit the emission of danger signals, which would otherwise favour bacterial clearance at the onset of infection. This work describes a novel regulatory mechanism of pore formation mediated by the T3SS, that are likely to be relevant for other extracellular pathogens.

Vyšlo v časopise: The Serine Protease EspC from Enteropathogenic Regulates Pore Formation and Cytotoxicity Mediated by the Type III Secretion System. PLoS Pathog 11(7): e32767. doi:10.1371/journal.ppat.1005013
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1005013

Souhrn

Zdroje

1. Campellone KG, Leong JM (2003) Tails of two Tirs: actin pedestal formation by enteropathogenic E. coli and enterohemorrhagic E. coli O157:H7. Curr Opin Microbiol 6: 82–90. 12615225

2. Schmidt MA (2010) LEEways: tales of EPEC, ATEC and EHEC. Cell Microbiol 12: 1544–1552. doi: 10.1111/j.1462-5822.2010.01518.x 20716205

3. Garmendia J, Frankel G, Crepin VF (2005) Enteropathogenic and enterohemorrhagic Escherichia coli infections: translocation, translocation, translocation. Infect Immun 73: 2573–2585. 15845459

4. Dautin N (2010) Serine Protease Autotransporters of Enterobacteriaceae (SPATEs): Biogenesis and Function. Toxins (Basel) 2: 1179–1206.

5. Dutta PR, Cappello R, Navarro-Garcia F, Nataro JP (2002) Functional comparison of serine protease autotransporters of enterobacteriaceae. Infect Immun 70: 7105–7113. 12438392

6. Drago-Serrano ME, Parra SG, Manjarrez-Hernandez HA (2006) EspC, an autotransporter protein secreted by enteropathogenic Escherichia coli (EPEC), displays protease activity on human hemoglobin. FEMS Microbiol Lett 265: 35–40. 17107418

7. Navarro-Garcia F, Serapio-Palacios A, Vidal JE, Salazar MI, Tapia-Pastrana G (2014) EspC promotes epithelial cell detachment by enteropathogenic Escherichia coli via sequential cleavages of a cytoskeletal protein and then focal adhesion proteins. Infect Immun 82: 2255–2265. doi: 10.1128/IAI.01386-13 24643541

8. Abreu AG, Bueris V, Porangaba TM, Sircili MP, Navarro-Garcia F, et al. (2013) Autotransporter protein-encoding genes of diarrheagenic Escherichia coli are found in both typical and atypical enteropathogenic E. coli strains. Appl Environ Microbiol 79: 411–414. doi: 10.1128/AEM.02635-12 23104414

9. Narimatsu H, Ogata K, Makino Y, Ito K (2010) Distribution of non-locus of enterocyte effacement pathogenic island-related genes in Escherichia coli carrying eae from patients with diarrhea and healthy individuals in Japan. J Clin Microbiol 48: 4107–4114. doi: 10.1128/JCM.00677-10 20844211

10. Elliott SJ, Sperandio V, Giron JA, Shin S, Mellies JL, et al. (2000) The locus of enterocyte effacement (LEE)-encoded regulator controls expression of both LEE- and non-LEE-encoded virulence factors in enteropathogenic and enterohemorrhagic Escherichia coli. Infect Immun 68: 6115–6126. 11035714

11. Mellies JL, Elliott SJ, Sperandio V, Donnenberg MS, Kaper JB (1999) The Per regulon of enteropathogenic Escherichia coli: identification of a regulatory cascade and a novel transcriptional activator, the locus of enterocyte effacement (LEE)-encoded regulator (Ler). Mol Microbiol 33: 296–306. 10411746

12. Vidal JE, Navarro-Garcia F (2006) Efficient translocation of EspC into epithelial cells depends on enteropathogenic Escherichia coli and host cell contact. Infect Immun 74: 2293–2303. 16552060

13. Vidal JE, Navarro-Garcia F (2008) EspC translocation into epithelial cells by enteropathogenic Escherichia coli requires a concerted participation of type V and III secretion systems. Cell Microbiol 10: 1975–1986. doi: 10.1111/j.1462-5822.2008.01181.x 18547338

14. Hartland EL, Daniell SJ, Delahay RM, Neves BC, Wallis T, et al. (2000) The type III protein translocation system of enteropathogenic Escherichia coli involves EspA-EspB protein interactions. Mol Microbiol 35: 1483–1492. 10760148

15. Ide T, Laarmann S, Greune L, Schillers H, Oberleithner H, et al. (2001) Characterization of translocation pores inserted into plasma membranes by type III-secreted Esp proteins of enteropathogenic Escherichia coli. Cell Microbiol 3: 669–679. 11580752

16. Nishimura K, Tajima N, Yoon YH, Park SY, Tame JR (2010) Autotransporter passenger proteins: virulence factors with common structural themes. J Mol Med (Berl) 88: 451–458.

17. Yen YT, Kostakioti M, Henderson IR, Stathopoulos C (2008) Common themes and variations in serine protease autotransporters. Trends Microbiol 16: 370–379. doi: 10.1016/j.tim.2008.05.003 18595714

18. Cornelis GR (2006) The type III secretion injectisome. Nat Rev Microbiol 4: 811–825. 17041629

19. Leverton LQ, Kaper JB (2005) Temporal expression of enteropathogenic Escherichia coli virulence genes in an in vitro model of infection. Infect Immun 73: 1034–1043. 15664947

20. Wachter C, Beinke C, Mattes M, Schmidt MA (1999) Insertion of EspD into epithelial target cell membranes by infecting enteropathogenic Escherichia coli. Mol Microbiol 31: 1695–1707. 10209743

21. Daniell SJ, Delahay RM, Shaw RK, Hartland EL, Pallen MJ, et al. (2001) Coiled-coil domain of enteropathogenic Escherichia coli type III secreted protein EspD is involved in EspA filament-mediated cell attachment and hemolysis. Infect Immun 69: 4055–4064. 11349076

22. Kresse AU, Rohde M, Guzman CA (1999) The EspD protein of enterohemorrhagic Escherichia coli is required for the formation of bacterial surface appendages and is incorporated in the cytoplasmic membranes of target cells. Infect Immun 67: 4834–4842. 10456938

23. Shaw RK, Daniell S, Ebel F, Frankel G, Knutton S (2001) EspA filament-mediated protein translocation into red blood cells. Cell Microbiol 3: 213–222. 11298645

24. Stein M, Kenny B, Stein MA, Finlay BB (1996) Characterization of EspC, a 110-kilodalton protein secreted by enteropathogenic Escherichia coli which is homologous to members of the immunoglobulin A protease-like family of secreted proteins. J Bacteriol 178: 6546–6554. 8932311

25. Clair C, Combettes L, Pierre F, Sansonetti P, Tran Van Nhieu G (2008) Extracellular-loop peptide antibodies reveal a predominant hemichannel organization of connexins in polarized intestinal cells. Exp Cell Res 314: 1250–1265. doi: 10.1016/j.yexcr.2007.12.021 18267319

26. Neyt C, Cornelis GR (1999) Insertion of a Yop translocation pore into the macrophage plasma membrane by Yersinia enterocolitica: requirement for translocators YopB and YopD, but not LcrG. Mol Microbiol 33: 971–981. 10476031

27. Gonzalez MR, Bischofberger M, Pernot L, van der Goot FG, Freche B (2008) Bacterial pore-forming toxins: the (w)hole story? Cell Mol Life Sci 65: 493–507. 17989920

28. Idone V, Tam C, Andrews NW (2008) Two-way traffic on the road to plasma membrane repair. Trends Cell Biol 18: 552–559. doi: 10.1016/j.tcb.2008.09.001 18848451

29. Luo W, Donnenberg MS (2011) Interactions and predicted host membrane topology of the enteropathogenic Escherichia coli translocator protein EspB. J Bacteriol 193: 2972–2980. doi: 10.1128/JB.00153-11 21498649

30. Ebel F, Podzadel T, Rohde M, Kresse AU, Kramer S, et al. (1998) Initial binding of Shiga toxin-producing Escherichia coli to host cells and subsequent induction of actin rearrangements depend on filamentous EspA-containing surface appendages. Mol Microbiol 30: 147–161. 9786192

31. Singh MP, Shaw RK, Knutton S, Pallen MJ, Crepin VF, et al. (2008) Identification of amino acid residues within the N-terminal domain of EspA that play a role in EspA filament biogenesis and function. J Bacteriol 190: 2221–2226. doi: 10.1128/JB.01753-07 18178741

32. Kenny B, Finlay BB (1995) Protein secretion by enteropathogenic Escherichia coli is essential for transducing signals to epithelial cells. Proc Natl Acad Sci U S A 92: 7991–7995. 7644526

33. Ochoa TJ, Noguera-Obenza M, Ebel F, Guzman CA, Gomez HF, et al. (2003) Lactoferrin impairs type III secretory system function in enteropathogenic Escherichia coli. Infect Immun 71: 5149–5155. 12933858

34. 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. Mol Cell Proteomics 11: 692–709. doi: 10.1074/mcp.M111.013672 22661456

35. Mellies JL, Navarro-Garcia F, Okeke I, Frederickson J, Nataro JP, et al. (2001) espC pathogenicity island of enteropathogenic Escherichia coli encodes an enterotoxin. Infect Immun 69: 315–324. 11119520

36. Navarro-Garcia F, Canizalez-Roman A, Sui BQ, Nataro JP, Azamar Y (2004) The serine protease motif of EspC from enteropathogenic Escherichia coli produces epithelial damage by a mechanism different from that of Pet toxin from enteroaggregative E. coli. Infect Immun 72: 3609–3621. 15155671

37. Navarro-Garcia F, Sonnested M, Teter K (2010) Host-Toxin Interactions Involving EspC and Pet, Two Serine Protease Autotransporters of the Enterobacteriaceae. Toxins (Basel) 2: 1134–1147.

38. Knodler LA, Vallance BA, Celli J, Winfree S, Hansen B, et al. (2010) Dissemination of invasive Salmonella via bacterial-induced extrusion of mucosal epithelia. Proc Natl Acad Sci U S A 107: 17733–17738. doi: 10.1073/pnas.1006098107 20876119

39. Vijayakumar V, Santiago A, Smith R, Smith M, Robins-Browne RM, et al. (2014) Role of class 1 serine protease autotransporter in the pathogenesis of Citrobacter rodentium colitis. Infect Immun 82: 2626–2636. doi: 10.1128/IAI.01518-13 24711562

40. Berger CN, Crepin VF, Baruch K, Mousnier A, Rosenshine I, et al. (2012) EspZ of enteropathogenic and enterohemorrhagic Escherichia coli regulates type III secretion system protein translocation. MBio 3.

41. Viboud GI, Bliska JB (2001) A bacterial type III secretion system inhibits actin polymerization to prevent pore formation in host cell membranes. Embo J 20: 5373–5382. 11574469

42. Roy K, Kansal R, Bartels SR, Hamilton DJ, Shaaban S, et al. (2011) Adhesin degradation accelerates delivery of heat-labile toxin by enterotoxigenic Escherichia coli. J Biol Chem 286: 29771–29779. doi: 10.1074/jbc.M111.251546 21757737

43. Brockmeyer J, Aldick T, Soltwisch J, Zhang W, Tarr PI, et al. (2011) Enterohaemorrhagic Escherichia coli haemolysin is cleaved and inactivated by serine protease EspPalpha. Environ Microbiol 13: 1327–1341. doi: 10.1111/j.1462-2920.2011.02431.x 21352460

44. Chantret I, Rodolosse A, Barbat A, Dussaulx E, Brot-Laroche E, et al. (1994) Differential expression of sucrase-isomaltase in clones isolated from early and late passages of the cell line Caco-2: evidence for glucose-dependent negative regulation. J Cell Sci 107 (Pt 1): 213–225. 8175910

45. Datsenko KA, Wanner BL (2000) One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. Proc Natl Acad Sci U S A 97: 6640–6645. 10829079