1887

Abstract

Type III secretion allows bacteria to inject effector proteins into host cells. In enteropathogenic (EPEC) the type III secreted protein, Tir, is translocated to the host-cell plasma membrane where it functions as a receptor for the bacterial adhesin intimin, leading to intimate bacterial attachment and ‘attaching and effacing’ (A/E) lesion formation. To study EPEC type III secretion the interaction of EPEC with monolayers of red blood cells (RBCs) has been exploited and in a recent study [Shaw, R. K., Daniell, S., Ebel, F., Frankel, G. & Knutton, S. (2001 ). 3, 213–222] it was shown that EPEC induced haemolysis of RBCs and translocation of EspD, a putative pore-forming type III secreted protein in the RBC membrane. Here it is demonstrated that EPEC are able to translocate and correctly insert Tir into the RBC membrane and produce an intimin–Tir intimate bacterial attachment, identical to that seen in A/E lesions. Following translocation Tir did not undergo any change in apparent molecular mass or become tyrosine-phosphorylated and there was no focusing of RBC cytoskeletal actin beneath intimately adherent bacteria, and no pedestal formation. This study, employing an RBC model of infection, has demonstrated that Tir translocation can be separated from host-cell-mediated Tir modifications; the data show that the EPEC type III protein translocation apparatus is sufficient to deliver and correctly insert Tir into host-cell membranes independent of eukaryotic cell functions.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-148-5-1355
2002-05-01
2019-10-19
Loading full text...

Full text loading...

/deliver/fulltext/micro/148/5/1481355a.html?itemId=/content/journal/micro/10.1099/00221287-148-5-1355&mimeType=html&fmt=ahah

References

  1. Blocker, A., Gounon, P., Larquet, E., Niebuhr, K., Cabiaux, V., Parsot, C. & Sansonetti, P. ( 1999; ). The tripartite type III secretion of Shigella flexneri inserts IpaB and IpaC into host membranes. J Cell Biol 147, 683-693.[CrossRef]
    [Google Scholar]
  2. Clerc, P., Baudry, B. & Sansonetti, P. J. ( 1986; ). Plasmid-mediated contact haemolytic activity in Shigella species: correlation with penetration into HeLa cells. Ann Inst Pasteur Microbiol 137A, 267-278.
    [Google Scholar]
  3. Donnenberg, M. S., Yu, J. & Kaper, J. B. ( 1993; ). A second chromosomal gene necessary for intimate attachment of enteropathogenic Escherichia coli to epithelial cells. J Bacteriol 175, 4670-4680.
    [Google Scholar]
  4. 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]
  5. 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.[CrossRef]
    [Google Scholar]
  6. Gauthier, A., de Grado, M. & Finlay, B. B. ( 2000; ). Mechanical fractionation reveals structural requirements for enteropathogenic Escherichia coli Tir insertion into host membranes. Infect Immun 68, 4344-4348.[CrossRef]
    [Google Scholar]
  7. Gomez-Duarte, O. G. & Kaper, J. B. ( 1995; ). A plasmid-encoded regulatory region activates chromosomal eaeA expression in enteropathogenic Escherichia coli. Infect Immun 63, 1767-1776.
    [Google Scholar]
  8. Goosney, D. L., DeVinney, R., Pfuetzner, R. A., Frey, E. A., Strynadka, N. C. & Finlay, B. B. ( 2000; ). Enteropathogenic E. coli translocated intimin receptor, Tir, interacts directly with alpha-actinin. Curr Biol 10, 735-738.[CrossRef]
    [Google Scholar]
  9. Hakansson, S., Schesser, K., Persson, C., Galyov, E. E., Rosqvist, R., Homble, F. & Wolf-Watz, H. ( 1996; ). The YopB protein of Yersinia pseudotuberculosis is essential for the translocation of Yop effector proteins across the target cell plasma membrane and displays a contact-dependent membrane disrupting activity. EMBO J 15, 5812-5823.
    [Google Scholar]
  10. 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.[CrossRef]
    [Google Scholar]
  11. 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]
  12. 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 USA 92, 7996-8000.[CrossRef]
    [Google Scholar]
  13. Jenkins, C., Chart, H., Smith, H. R., Hartland, E. L., Batchelor, M., Delahay, R. M., Dougan, G. & Frankel, G. ( 2000; ). Antibody response of patients infected with verocytotoxin-producing Escherichia coli to protein antigens encoded on the LEE locus. J Med Microbiol 49, 97-101.
    [Google Scholar]
  14. Jerse, A. E., Yu, J., Tall, B. D. & Kaper, J. B. ( 1990; ). A genetic locus of enteropathogenic Escherichia coli necessary for the production of attaching and effacing lesions on tissue culture cells. Proc Natl Acad Sci USA 87, 7839-7843.[CrossRef]
    [Google Scholar]
  15. Kenny, B. ( 1999; ). Phosphorylation of tyrosine 474 of the enteropathogenic Escherichia coli (EPEC) Tir receptor molecule is essential for actin nucleating activity and is preceded by additional host modifications. Mol Microbiol 31, 1229-1241.[CrossRef]
    [Google Scholar]
  16. Kenny, B. & Finlay, B. B. ( 1997; ). Intimin-dependent binding of enteropathogenic Escherichia coli to host cells triggers novel signaling events, including tyrosine phosphorylation of phospholipase C-gamma1. Infect Immun 65, 2528-2536.
    [Google Scholar]
  17. Kenny, B. & Jepson, M. ( 2000; ). Targeting of an enteropathogenic E. coli (EPEC) effector protein to host mitochondria. Cell Microbiol 2, 579-590.[CrossRef]
    [Google Scholar]
  18. 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.[CrossRef]
    [Google Scholar]
  19. Kenny, B., DeVinney, R., Stein, M., Reinscheid, D. J., Frey, E. A. & Finlay, B. B. ( 1997; ). Enteropathogenic E. coli (EPEC) transfers its receptor for intimate adherence into mammalian cells. Cell 91, 511-520.[CrossRef]
    [Google Scholar]
  20. Knutton, S. ( 1995; ). Electron microscopical methods in adhesion. Methods Enzymol 253, 145-158.
    [Google Scholar]
  21. Knutton, S., Lloyd, D. R. & McNeish, A. S. ( 1987; ). Adhesion of enteropathogenic Escherichia coli to human intestinal enterocytes and cultured human intestinal mucosa. Infect Immun 55, 69-77.
    [Google Scholar]
  22. 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]
  23. Knutton, S., Adu-Bobie, J., Bain, C., Phillips, A. D., Dougan, G. & Frankel, G. ( 1997; ). Down regulation of intimin expression during attaching and effacing enteropathogenic Escherichia coli adhesion. Infect Immun 65, 1644-1652.
    [Google Scholar]
  24. 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.[CrossRef]
    [Google Scholar]
  25. 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]
  26. 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]
  27. Li, Y., Frey, E., Mackenzie, A. M. R. & Finlay, B. B. ( 2000; ). Human response to Escherichia coli O157:H7 infection: antibodies to secreted virulence factors. Infect Immun 68, 5090-5095.[CrossRef]
    [Google Scholar]
  28. McDaniel, T. K. & Kaper, J. B. ( 1997; ). A cloned pathogenicity island from enteropathogenic Escherichia coli confers the attaching and effacing phenotype on E. coli K-12. Mol Microbiol 23, 399-407.[CrossRef]
    [Google Scholar]
  29. McNamara, B. P., Koutsouris, A., O’Connell, C. B., Nougayrede, J. P., Donnenberg, M. S. & Hecht, G. ( 2001; ). Translocated EspF protein from enteropathogenic Escherichia coli disrupts host intestinal barrier function. J Clin Invest 107, 621-629.[CrossRef]
    [Google Scholar]
  30. Mellies, J. L., Elliott, S. J., Sperandio, V., Donnenberg, M. S. & Kaper, J. B. ( 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.[CrossRef]
    [Google Scholar]
  31. Rosenshine, I., Donnenberg, M. S., Kaper, J. B. & Finlay, B. B. ( 1992; ). Signal transduction between enteropathogenic Escherichia coli (EPEC) and epithelial cells: EPEC induces tyrosine phosphorylation of host cell proteins to initiate cytoskeletal rearrangement and bacterial uptake. EMBO J 11, 3551-3560.
    [Google Scholar]
  32. Rosenshine, I., Ruschkowski, S., Stein, M., Reinscheid, D. J., Mills, S. D. & Finlay, B. B. ( 1996; ). A pathogenic bacterium triggers epithelial signals to form a functional bacterial receptor that mediates actin pseudopod formation. EMBO J 15, 2613-2624.
    [Google Scholar]
  33. 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.[CrossRef]
    [Google Scholar]
  34. Wachter, C., Beinke, C., Mattes, M. & Schmidt, M. A. ( 1999; ). Insertion of EspD into epithelial target cell membranes by infecting enteropathogenic Escherichia coli. Mol Microbiol 31, 1695-1707.[CrossRef]
    [Google Scholar]
  35. Warawa, J. & Kenny, B. ( 2001; ). Phosphoserine modification of the enteropathogenic Escherichia coli Tir molecule is required to trigger conformational changes in Tir and efficient pedestal elongation. Mol Microbiol 42, 1269-1280.
    [Google Scholar]
  36. Warawa, J., Finlay, B. B. & Kenny, B. ( 1999; ). Type III secretion-dependent hemolytic activity of enteropathogenic Escherichia coli. Infect Immun 67, 5538-5540.
    [Google Scholar]
  37. Wolff, C., Nisan, I., Hanski, E., Frankel, G. & Rosenshine, I. ( 1998; ). Protein translocation into HeLa cells by infecting enteropathogenic Escherichia coli. Mol Microbiol 28, 143-155.
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-148-5-1355
Loading
/content/journal/micro/10.1099/00221287-148-5-1355
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error