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Volume 149, Issue 12

CesAB is an enteropathogenic chaperone for the type-III translocator proteins EspA and EspB Free

Abstract

Enteropathogenic (EPEC) are extracellular pathogens that colonize mucosal surfaces of the intestine via formation of attaching and effacing (A/E) lesions. The genes responsible for induction of the A/E lesions are located on a pathogenicity island, termed the locus of enterocyte effacement (LEE), which encodes the adhesin intimin and the type III secretion system needle complex, translocator and effector proteins. One of the major EPEC translocator proteins, EspA, forms a filamentous conduit along which secreted proteins travel before they arrive at the translocation pore in the plasma membrane of the host cell, which is composed of EspB and EspD. Prior to secretion, many type III proteins, including translocators, are maintained in the bacterial cytoplasm by association with a specific chaperone. In EPEC, chaperones have been identified for the effector proteins Tir, Map and EspF, and the translocator proteins EspD and EspB. In this study, CesAB (Orf3 of the LEE) was identified as a chaperone for EspA and EspB. Specific CesAB–EspA and CesAB–EspB protein interactions are demonstrated. CesAB was essential for stability of EspA within the bacterial cell prior to secretion. Furthermore, a mutant failed to secrete EspA, as well as EspB, to assemble EspA filaments, to induce A/E lesion following infection of HEp-2 cells and to adhere to, or cause haemolysis of, erythrocytes.

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
Keyword(s): A/E, attaching and effacing , EPEC, enteropathogenic Escherichia coli , FAS, fluorescence actin staining , LEE, locus of enterocyte effacement , RBC, red blood cells and TTSS, type III secretion system
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2003-12-01
2024-04-16
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References

  1. Abe A., de Grado M., Pfuetzner R. A., Sanchez-SanMartin C., DeVinney R., Puente J. L., Strynadka N. C. J., Finlay B. B. 1999; Enteropathogenic Escherichia coli translocated intimin receptor, Tir, requires a specific chaperone for stable secretion. Mol Microbiol 33:1162–1175
     [Google Scholar]
  2. Anderson D. M., Ramamurthi K. S., Tam C., Schneewind O. 2002; YopD and LcrH regulate expression of Yersinia enterocolitica YopQ by a posttranscriptional mechanism and bind to yopQ RNA. J Bacteriol 184:1287–1295
     [Google Scholar]
  3. Blocker A., Jouihri N., Larquet E., Gounon P., Ebel F., Parsot C., Sansonetti P., Allaoui A. 2001; Structure and composition of the Shigella flexneri ‘needle complex’, a part of its type III secreton. Mol Microbiol 39:652–663
     [Google Scholar]
  4. Boyd A. P., Lambermont I., Cornelis G. R. 2000; Competition between the Yops of Yersinia enterocolitica for delivery into eukaryotic cells: role of the SycE chaperone binding domain of YopE. J Bacteriol 182:4811–4821
     [Google Scholar]
  5. Cambronne E. D., Cheng L. W., Schneewind O. 2000; LcrQ/YscM1, regulators of the Yersinia yop virulon, are injected into host cells by a chaperone-dependent mechanism. Mol Microbiol 37:263–273
     [Google Scholar]
  6. Creasey E. A., Delahay R. M., Bishop A. A., Shaw R. K., Kenny B., Knutton S., Frankel G. 2003a; CesT is a bivalent enteropathogenic Escherichia coli chaperone required for translocation of both Tir and Map. Mol Microbiol 47:209–221
     [Google Scholar]
  7. Creasey E. A., Delahay R. M., Daniell S. J., Frankel G. 2003b; Yeast two-hybrid system survey of interactions between LEE-encoded proteins of enteropathogenic Escherichia coli . Microbiology 149:2093–2106
     [Google Scholar]
  8. Daniell S. J., Takahashi N., Wilson R. 7 other authors 2001; The filamentous type III secretion translocon of enteropathogenic Escherichia coli . Cell Microbiol 3:865–871
     [Google Scholar]
  9. Daniell S. J., Kocsis E., Morris E., Knutton S., Booy F. P., Frankel G. 2003; 3D structure of EspA filaments from enteropathogenic Escherichia coli . Mol Microbiol 49:301–308
     [Google Scholar]
  10. Darwin K. H., Miller V. L. 2001; Type III secretion chaperone-dependent regulation: activation of virulence genes by SicA and InvF in Salmonella typhimurium . EMBO J 20:1850–1862
     [Google Scholar]
  11. Datsenko K. A., Wanner B. L. 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
     [Google Scholar]
  12. Day J. B., Plano G. V. 1998; A complex composed of SycN and YscB functions as a specific chaperone for YopN in Yersinia pestis . Mol Microbiol 30:777–788
     [Google Scholar]
  13. Delahay R. M., Knutton S., Shaw R. K., Hartland E. L., Pallen M. J., Frankel G. 1999; The coiled-coil domain of EspA is essential for the assembly of the type III secretion translocon on the surface of enteropathogenic Escherichia coli . J Biol Chem 274:35969–35974
     [Google Scholar]
  14. Deng W., Li Y., Vallance B. A., Finlay B. B. 2001; Locus of enterocyte effacement from Citrobacter rodentium : sequence analysis and evidence for horizontal transfer among attaching and effacing pathogens. Infect Immun 69:6323–6335
     [Google Scholar]
  15. Ebel F., Podzadel T., Rohde M., Kresse A. U., Kramer S., Deibel C., Guzman C. A., Chakraborty T. 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
     [Google Scholar]
  16. Elliott S. J., Wainwright L. A., McDaniel T. K., Jarvis K. G., Deng Y., Lai L. C., McNamara B. P., Donnenberg M. S., Kaper J. B. 1998; The complete sequence of the locus of enterocyte effacement (LEE) from enteropathogenic Escherichia coli E2348/69. Mol Microbiol 28:1–4
     [Google Scholar]
  17. Elliott S. J., Hutcheson S. W., Dubois M. S., Mellies J. L., Wainwright L. A., Batchelor M., Frankel G., Knutton S., Kaper J. B. 1999; Identification of CesT, a chaperone for the type III secretion of Tir in enteropathogenic Escherichia coli . Mol Microbiol 33:1176–1189
     [Google Scholar]
  18. Elliott S. J., O'Connell C. B., Koutsouris A., Brinkley C., Donnenberg M. S., Hecht G., Kaper J. B. 2002; A gene from the locus of enterocyte effacement that is required for enteropathogenic Escherichia coli to increase tight-junction permeability encodes a chaperone for EspF. Infect Immun 70:2271–2277
     [Google Scholar]
  19. Francis M. S., Aili M., Wiklund M. L., Wolf-Watz H. 2000; A study of the YopD-LcrH interaction from Yersinia pseudotuberculosis reveals a role for hydrophobic residues within the amphipathic domain of YopD. Mol Microbiol 38:85–102
     [Google Scholar]
  20. Frankel G., Phillips A. D., Rosenshine I., Dougan G., Kaper J. B., Knutton S. 1998; Enteropathogenic and enterohaemorrhagic Escherichia coli : more subversive elements. Mol Microbiol 30:911–921
     [Google Scholar]
  21. Friedberg D., Umanski T., Fang Y., Rosenshine I. 1999; Hierarchy in the expression of the locus of enterocyte effacement genes of enteropathogenic Escherichia coli . Mol Microbiol 34:941–952
     [Google Scholar]
  22. Frithz-Lindsten E., Rosqvist R., Johansson L., Forsberg A. 1995; The chaperone-like protein YerA of Yersinia pseudotuberculosis stabilises YopE in the cytoplasm but is dispensable for targetting to the secretion loci. Mol Microbiol 16:635–647
     [Google Scholar]
  23. Fu Y., Galan J. E. 1998; Identification of a specific chaperone for SptP, a substrate of the centisome 63 type III secretion system. J Bacteriol 180:3393–3399
     [Google Scholar]
  24. Hartland E. L., Robins-Browne R. M. 1998; In vitro association between the virulence proteins, YopD and YopE, of Yersinia enterocolitica . FEMS Microbiol Lett 162:207–213
     [Google Scholar]
  25. Hartland E. L., Batchelor M., Delahay R. M., Hale C., Matthews S., Dougan G., Knutton S., Connerton I., Frankel G. 1999; Binding of intimin from enteropathogenic Escherichia coli to Tir and to host cells. Mol Microbiol 32:151–158
     [Google Scholar]
  26. Hartland E. L., Daniell S. J., Delahay R. M., Neves B. C., Wallis T., Shaw R. K., Hale C., Knutton S., Frankel G. 2000; The type III protein translocation system of enteropathogenic Escherichia coli involves EspA-EspB protein interactions. Mol Microbiol 35:1483–1492
     [Google Scholar]
  27. Hueck C. J. 1998; Type III protein secretion systems in bacterial pathogens of animals and plants. Microbiol Mol Biol Rev 62:379–433
     [Google Scholar]
  28. Jackson M. W., Day J. B., Plano G. V. 1998; YscB of Yersinia pestis functions as a specific chaperone for YopN. J Bacteriol 180:4912–4921
     [Google Scholar]
  29. Jarvis K. G., Giron J. A., Jerse A. E., McDaniel T. K., Donnenberg M. S., Kaper J. B. 1995; Enteropathogenic Escherichia coli contains a putative type III secretion system necessary for the export of proteins involved in attaching and effacing lesion formation. Proc Natl Acad Sci U S A 92:7996–8000
     [Google Scholar]
  30. Kenny B., Lai L. C., Finlay B. B., Donnenberg M. S. 1996; EspA, a protein secreted by enteropathogenic Escherichia coli , is required to induce signals in epithelial cells. Mol Microbiol 20:313–323
     [Google Scholar]
  31. Kenny B., Abe A., Stein M., Finlay B. B. 1997; Enteropathogenic Escherichia coli protein secretion is induced in response to conditions similar to those in the gastrointestinal tract. Infect Immun 65:2606–2612
     [Google Scholar]
  32. Knutton S., Baldwin T., Williams P. H., McNeish A. S. 1989; Actin accumulation at sites of bacterial adhesion to tissue culture cells – basis of a new diagnostic test for enteropathogenic and enterohemorrhagic Escherichia coli . Infect Immun 57:1290–1298
     [Google Scholar]
  33. Knutton S., Rosenshine I., Pallen M. J., Nisan I., Neves B. C., Bain C., Wolff C., Dougan G., Frankel G. 1998; A novel EspA-associated surface organelle of enteropathogenic Escherichia coli involved in protein translocation into epithelial cells. EMBO J 17:2166–2176
     [Google Scholar]
  34. Knutton S., Shaw R., Frankel G. 2002; Interaction of enteropathogenic Escherichia coli with red blood cell monolayers. Methods Enzymol 358:350–355
     [Google Scholar]
  35. Kubori T., Matsushima Y., Nakamura D., Uralil J., Lara-Tejero M., Sukhan A., Galan J. E., Aizawa S. I. 1998; Supramolecular structure of the Salmonella typhimurium type III protein secretion system. Science 280:602–605
     [Google Scholar]
  36. Lai L. C., Wainwright L. A., Stone K. D., Donnenberg M. S. 1997; A third secreted protein that is encoded by the enteropathogenic Escherichia coli pathogenicity island is required for transduction of signals and for attaching and effacing activities in host cells. Infect Immun 65:2211–2217
     [Google Scholar]
  37. Levine M. M., Bergquist E. J., Nalin D. R., Waterman D. H., Hornick R. B., Young C. R., Sotman S. 1978; Escherichia coli strains that cause diarrhoea but do not produce heat-labile or heat-stable enterotoxins and are non-invasive. Lancet 1:1119–1122
     [Google Scholar]
  38. Mavris M., Page A. L., Tournebize R., Demers B., Sansonetti P., Parsot C. 2002; Regulation of transcription by the activity of the Shigella flexneri type III secretion apparatus. Mol Microbiol 43:1543–1553
     [Google Scholar]
  39. McDaniel T. K., Jarvis K. G., Donnenberg M. S., Kaper J. B. 1995; A genetic locus of enterocyte effacement conserved among diverse enterobacterial pathogens. Proc Natl Acad Sci U S A 92:1664–1668
     [Google Scholar]
  40. Menard R., Sansonetti P., Parsot C., Vasselon T. 1994; Extracellular association and cytoplasmic partitioning of the IpaB and IpaC invasins of S. flexneri . Cell 79:515–525
     [Google Scholar]
  41. Neves B. C., Mundy R., Petrovska L., Dougan G., Knutton S., Frankel G. 2003; CesD2 of enteropathogenic Escherichia coli is a second chaperone for the type III secretion translocator protein EspD. Infect Immun 71:2130–2141
     [Google Scholar]
  42. Neyt C., Cornelis G. R. 1999; Role of SycD, the chaperone of the Yersinia Yop translocators YopB and YopD. Mol Microbiol 31:143–156
     [Google Scholar]
  43. Niebuhr K., Jouihri N., Allaoui A., Gounon P., Sansonetti P. J., Parsot C. 2000; IpgD, a protein secreted by the type III secretion machinery of Shigella flexneri , is chaperoned by IpgE and implicated in entry focus formation. Mol Microbiol 38:8–19
     [Google Scholar]
  44. Page A. L., Parsot C. 2002; Chaperones of the type III secretion pathway: jacks of all trades. Mol Microbiol 46:1–11
     [Google Scholar]
  45. Page A. L., Sansonetti P., Parsot C. 2002; Spa15 of Shigella flexneri , a third type of chaperone in the type III secretion pathway. Mol Microbiol 43:1533–1542
     [Google Scholar]
  46. Ruiz-Albert J., Mundy R., Yu X. J., Beuzon C. R., Holden D. W. 2003; SseA is a chaperone for the SseB and SseD translocon components of the Salmonella pathogenicity-island-2-encoded type III secretion system. Microbiology 149:1103–1111
     [Google Scholar]
  47. Schlosser-Silverman E., Elgrably-Weiss M., Rosenshine I., Kohen R., Altuvia S. 2000; Characterization of Escherichia coli DNA lesions generated within J774 macrophages. J Bacteriol 182:5225–5230
     [Google Scholar]
  48. Sekiya K., Ohishi M., Ogino T., Tamano K., Sasakawa C., Abe A. 2001; Supermolecular structure of the enteropathogenic Escherichia coli type III secretion system and its direct interaction with the EspA-sheath-like structure. Proc Natl Acad Sci U S A 98:11638–11643
     [Google Scholar]
  49. Shaw R. K., Daniell S., Ebel F., Frankel G., Knutton S. 2001; EspA filament-mediated protein translocation into red blood cells. Cell Microbiol 3:213–222
     [Google Scholar]
  50. Tamano K., Aizawa S. I., Katayama E., Nonaka T., Imajoh-Ohmi S., Kuwae A., Nagai S., Sasakawa C. 2000; Supramolecular structure of the Shigella type III secretion machinery: the needle part is changeable in length and essential for delivery of effectors. EMBO J 19:3876–3887
     [Google Scholar]
  51. Tauschek M., Strugnell R. A., Robins-Browne R. M. 2002; Characterization and evidence of mobilization of the LEE pathogenicity island of rabbit-specific strains of enteropathogenic Escherichia coli . Mol Microbiol 44:1533–1550
     [Google Scholar]
  52. Tucker S. C., Galan J. E. 2000; Complex function for SicA, a Salmonella enterica serovar Typhimurium type III secretion-associated chaperone. J Bacteriol 182:2262–2268
     [Google Scholar]
  53. Umanski T., Rosenshine I., Friedberg D. 2002; Thermoregulated expression of virulence genes in enteropathogenic Escherichia coli . Microbiology 148:2735–2744
     [Google Scholar]
  54. Wainwright L. A., Kaper J. B. 1998; EspB and EspD require a specific chaperone for proper secretion from enteropathogenic Escherichia coli . Mol Microbiol 27:1247–1260
     [Google Scholar]
  55. Wattiau P., Bernier B., Deslee P., Michiels T., Cornelis G. R. 1994; Individual chaperones required for Yop secretion by Yersinia . Proc Natl Acad Sci U S A 91:10493–10497
     [Google Scholar]
  56. Wilson R. K., Shaw R. K., Daniell S., Knutton S., Frankel G. 2001; Role of EscF, a putative needle complex protein, in the type III protein translocation system of enteropathogenic Escherichia coli . Cell Microbiol 3:753–762
     [Google Scholar]
  57. Woestyn S., Sory M. P., Boland A., Lequenne O., Cornelis G. R. 1996; The cytosolic SycE and SycH chaperones of Yersinia protect the region of YopE and YopH involved in translocation across eukaryotic cell membranes. Mol Microbiol 20:1261–1271
     [Google Scholar]
  58. Wulff-Strobel C. R., Williams A. W., Straley S. C. 2002; LcrQ and SycH function together at the Ysc type III secretion system in Yersinia pestis to impose a hierarchy of secretion. Mol Microbiol 43:411–423
     [Google Scholar]
  59. Zhu C., Agin T. S., Elliott S. J., Johnson L. A., Thate T. E., Kaper J. B., Boedeker E. C. 2001; Complete nucleotide sequence and analysis of the locus of enterocyte effacement from rabbit diarrheagenic Escherichia coli RDEC-1. Infect Immun 69:2107–2115
     [Google Scholar]
  60. Zurawski D. V., Stein M. A. 2003; SseA acts as the chaperone for the SseB component of the Salmonella Pathogenicity Island 2 translocon. Mol Microbiol 47:1341–1351
     [Google Scholar]
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CesAB is an enteropathogenic Escherichia coli chaperone for the type-III translocator proteins EspA and EspB
Microbiology 149, 3639 (2003); https://doi.org/10.1099/mic.0.26735-0
/content/journal/micro/10.1099/mic.0.26735-0
/content/journal/micro/10.1099/mic.0.26735-0
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