1887

Abstract

The major secreted virulence factor of , the vacuolating cytotoxin VacA, is known to insert into eukaryotic membranes and has been observed in association with the surface of cells that are actively producing it. Here, it is demonstrated that VacA is capable of interacting with the surface of and after secretion. It is shown that this interaction is resistant to disruption of electrostatic and hydrophobic forces, and that it appears to occur despite truncation of LPS and the removal of trypsin-accessible surface proteins. Adsorption to bacterial cell surfaces was independent of the VacA subtype, suggesting that it is not mediated through recognition of a known receptor by the VacA p58 subunit. Similarly, adsorption to bacterial cell surfaces is unlikely to be instigated by the extreme N-terminus of VacA, since a hydrophilic extension at this location that is known to disrupt VacA-induced vacuolation did not interfere with adsorption to cells.

Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.45946-0
2005-07-01
2019-11-18
Loading full text...

Full text loading...

/deliver/fulltext/jmm/54/7/JM540702.html?itemId=/content/journal/jmm/10.1099/jmm.0.45946-0&mimeType=html&fmt=ahah

References

  1. Adrian, M., Cover, T. L., Dubochet, J. & Heuser, J. E. ( 2002;). Multiple oligomeric states of the Helicobacter pylori vacuolating toxin demonstrated by cryo-electron microscopy. J Mol Biol 318, 121–133.[CrossRef]
    [Google Scholar]
  2. Atherton, J. C., Cao, P., Peek, R. M., Jr, Tummuru, M. K. R., Blaser, M. J. & Cover, T. L. ( 1995;). Mosaicism in vacuolating cytotoxin alleles of Helicobacter pylori.Association of specific vacA types with cytotoxin production and peptic ulceration. J Biol Chem 270, 17771–17777.[CrossRef]
    [Google Scholar]
  3. Bebb, J. R., Letley, D. P., Rhead, J. L. & Atherton, J. C. ( 2003;). Helicobacter pylori supernatants cause epithelial cytoskeletal disruption that is bacterial strain and epithelial cell line dependent, but not toxin VacA dependent. Infect Immun 71, 3623–3627.[CrossRef]
    [Google Scholar]
  4. Boman, H. G. & Monner, D. A. ( 1975;). Characterization of lipopolysaccharides from Escherichia coli K-12 mutants. J Bacteriol 121, 455–464.
    [Google Scholar]
  5. Boman, H. G., Eriksson-Grennberg, K. G., Normark, S. & Matsson, E. ( 1968;). Resistance of Escherichia coli to penicillins.IV. Genetic study of mutants resistant to d,l-ampicillin concentrations of 100 μg ml−1. Genet Res 12, 169–185.[CrossRef]
    [Google Scholar]
  6. Casadaban, M. J. ( 1976;). Transposition and fusion of the lac genes to selected promoters in Escherichia coli using bacteriophage lambda and Mu. J Mol Biol 104, 541–555.[CrossRef]
    [Google Scholar]
  7. Cover, T. L. ( 1996;). The vacuolating cytotoxin of Helicobacter pylori. Mol Microbiol 20, 241–246.[CrossRef]
    [Google Scholar]
  8. Cover, T. L. & Blaser, M. J. ( 1992;). Purification and characterization of the vacuolating toxin from Helicobacter pylori. J Biol Chem 267, 10570–10575.
    [Google Scholar]
  9. Cover, T. L., Hanson, P. I. & Heuser, J. E. ( 1997;). Acid induced dissociation of VacA, the Heliocbacter pylori vacuolating cytotoxin, reveals its pattern of assembly. J Cell Biol 138, 759–769.[CrossRef]
    [Google Scholar]
  10. Czajkowsky, D. M., Iwamoto, H., Cover, T. L. & Shao, Z. ( 1999;). The vacuolating toxin from Helicobacter pylori forms hexameric pores in lipid bilayers at low pH. Proc Natl Acad Sci U S A 96, 2001–2006.[CrossRef]
    [Google Scholar]
  11. Edwards, N. J., Monteiro, M. A., Faller, G., Walsh, E. J., Moran, A. P., Roberts, I. S. & High, N. J. ( 2000;). Lewis X structures in the O antigen side-chain promote adhesion of Helicobacter pylori to the gastric epithelium. Mol Microbiol 35, 1530–1539.
    [Google Scholar]
  12. Ernst, P. B. & Gold, B. D. ( 2000;). The disease spectrum of Helicobacter pylori: the immunopathogenesis of gastroduodenal ulcer and gastric cancer. Annu Rev Microbiol 54, 615–640.[CrossRef]
    [Google Scholar]
  13. Fiocca, R., Necchi, V., Sommi, P., Ricci, V., Telford, J., Cover, T. L. & Solcia, E. ( 1999;). Release of Helicobacter pylori vacuolating cytotoxin by both a specific secretion pathway and budding of outer membrane vesicles.Uptake of released toxin and vesicles by gastric epithelium. J Pathol 188, 220–226.[CrossRef]
    [Google Scholar]
  14. Fitchen, N., O'Shea, P., Williams, P. & Hardie, K. R. ( 2003;). Electrostatic sensor for identifying interactions between peptides and bacterial membranes. Mol Immunol 40, 407–411.[CrossRef]
    [Google Scholar]
  15. Fujikawa, A., Shirasaka, D., Yamamoto, S. & 9 other authors ( 2003;). Mice deficient in protein tyrosine phosphatase receptor type Z are resistant to gastric ulcer induction by VacA of Helicobacter pylori. Nat Genet 33, 375–381.[CrossRef]
    [Google Scholar]
  16. Galmiche, A., Rassow, J., Doye, A. & 9 other authors ( 2000;). The N-terminal 34 kDa fragment of Helicobacter pylori vacuolating cytotoxin targets mitochondria and induces cytochrome c release. EMBO J 19, 6361–6370.[CrossRef]
    [Google Scholar]
  17. Garner, J. A. & Cover, T. L. ( 1996;). Binding and internalization of the Helicobacter pylori vacuolating cytotoxin by epithelial cells. Infect Immun 64, 4197–4203.
    [Google Scholar]
  18. Gauthier, N. C., Ricci, V., Gounon, P., Doye, A., Tauc, M., Poujeol, P. & Boquet, P. ( 2004;). Glycosylphosphatidylinositol-anchored proteins and actin cytoskeleton modulate chloride transport by channels formed by the Helicobacter pylori vacuolating cytotoxin VacA in HeLa cells. J Biol Chem 279, 9481–9489.[CrossRef]
    [Google Scholar]
  19. Gebert, B., Fischer, W. & Haas, R. ( 2004;). The Helicobacter pylori vacuolating cytotoxin: from cellular vacuolation to immunosuppressive activities. Rev Physiol Biochem Pharmacol 152, 205–220.
    [Google Scholar]
  20. Geisse, N. A., Cover, T. L., Henderson, R. M. & Edwardson, J. M. ( 2004;). Targeting of Helicobacter pylori vacuolating toxin to lipid raft membrane domains analysed by atomic force microscopy. Biochem J 381, 911–917.[CrossRef]
    [Google Scholar]
  21. Hardie, K. R., Cooksley, C., Green, A. D. & Winzer, K. ( 2003;). Autoinducer 2 activity in Escherichia coli culture supernatants can be actively reduced despite maintenance of an active synthase, LuxS. Microbiology 149, 715–728.[CrossRef]
    [Google Scholar]
  22. Henderson, I. R., Navarro-Garcia, F. & Nataro, J. P. ( 1998;). The great escape: structure and function of the autotransporter proteins. Trends Microbiol 6, 370–378.[CrossRef]
    [Google Scholar]
  23. Henderson, I. R., Cappello, R. & Nataro, J. P. ( 2000;). Autotransporter proteins, evolution and redefining protein secretion. Trends Microbiol 8, 529–532.[CrossRef]
    [Google Scholar]
  24. Ilver, D., Barone, S., Mercati, D., Lupetti, P. & Telford, J. L. ( 2004;). Helicobacter pylori toxin VacA is transferred to host cells via a novel contact-dependent mechanism. Cell Microbiol 6, 167–174.[CrossRef]
    [Google Scholar]
  25. Ji, X., Fernandez, T., Burroni, D., Pagliaccia, C., Atherton, J. C., Reyrat, J. M., Rappuoli, R. & Telford, J. L. ( 2000;). Cell specificity of Helicobacter pylori cytotoxin is determined by a short region in the polymorphic midregion. Infect Immun 68, 3754–3757.[CrossRef]
    [Google Scholar]
  26. Keenan, J., Day, T., Neal, S., Cook, B., Perez-Perez, G., Allardyce, R. & Bagshaw, P. ( 2000;). A role for the bacterial outer membrane in the pathogenesis of Helicobacter pylori infection. FEMS Microbiol Lett 182, 259–264.[CrossRef]
    [Google Scholar]
  27. Kidd, M., Lastovica, A. J., Atherton, J. C. & Louw, J. A. ( 1999;). Heterogeneity in the Helicobacter pylori vacA and cagA genes: association with gastroduodenal disease in South Africa? Gut 45, 499–502.[CrossRef]
    [Google Scholar]
  28. Kim, S., Chamberlain, A. K. & Bowie, J. U. ( 2004;). Membrane channel structure of Helicobacter pylori vacuolating toxin: role of multiple GXXXG motifs in cylindrical channels. Proc Natl Acad Sci U S A 101, 5988–5991.[CrossRef]
    [Google Scholar]
  29. Kimura, M., Goto, S., Wada, A., Yahiro, K., Niidome, T., Hatakeyama, T., Aoyagi, H., Hirayama, T. & Kondo, T. ( 1999;). Vacuolating cytotoxin purified from Helicobacter pylori causes mitochondrial damage in human gastric cells. Microb Pathog 26, 45–52.[CrossRef]
    [Google Scholar]
  30. Kuck, D., Kolmerer, B., Iking-Konert, C., Krammer, P. H., Stremmel, W. & Rudi, J. ( 2001;). Vacuolating cytotoxin of Helicobacter pylori induces apoptosis in the human gastric epithelial cell line AGS. Infect Immun 69, 5080–5087.[CrossRef]
    [Google Scholar]
  31. Kukkonen, M. & Korhonen, T. K. ( 2004;). The omptin family of enterobacterial surface proteases/adhesins: from housekeeping in Escherichia coli to systemic spread of Yersinia pestis. Int J Med Microbiol 294, 7–14.[CrossRef]
    [Google Scholar]
  32. Letley, D. P. & Atherton, J. C. ( 2000;). Natural diversity in the N terminus of the mature vacuolating cytotoxin of Helicobacter pylori determines cytotoxin activity. J Bacteriol 182, 3278–3280.[CrossRef]
    [Google Scholar]
  33. Letley, D. P., Lastovica, A., Louw, J. A., Hawkey, C. J. & Atherton, J. C. ( 1999;). Allelic diversity of the Helicobacter pylori vacuolating cytotoxin gene in South Africa: rarity of the vacA s1a genotype and natural occurrence of an s2/m1 allele. J Clin Microbiol 37, 1203–1205.
    [Google Scholar]
  34. Letley, D. P., Rhead, J. L., Twells, R. J., Dove, B. & Atherton, J. C. ( 2003;). Determinants of non-toxicity in the gastric pathogen Helicobacter pylori. J Biol Chem 278, 26734–26741.[CrossRef]
    [Google Scholar]
  35. Leunk, R. D., Johnson, P. T., David, B. C., Kraft, W. G. & Morgan, D. R. ( 1988;). Cytotoxic activity in broth-culture filtrates of Campylobacter pylori. J Med Microbiol 26, 93–99.[CrossRef]
    [Google Scholar]
  36. Li, Y., Wandinger-Ness, A., Goldenring, J. R. & Cover, T. L. ( 2004;). Clustering and redistribution of late endocytic compartments in response to Helicobacter pylori vacuolating toxin. Mol Biol Cell 15, 1946–1959.[CrossRef]
    [Google Scholar]
  37. Lupetti, P., Heuser, J. E., Manetti, R., Massari, P., Lanzavecchia, S., Bellon, P. L., Dallai, R., Rappuoli, R. & Telford, J. L. ( 1996;). Oligomeric and subunit structure of the Helicobacter pylori vacuolating cytotoxin. J Cell Biol 133, 801–807.[CrossRef]
    [Google Scholar]
  38. McClain, M. S., Iwamoto, H., Cao, P., Vinion-Dubiel, A. D., Li, Y., Szabo, G., Shao, Z. & Cover, T. L. ( 2003;). Essential role of a GXXXG motif for membrane channel formation by Helicobacter pylori vacuolating toxin. J Biol Chem 278, 12101–12108.[CrossRef]
    [Google Scholar]
  39. Menaker, R. J., Ceponis, P. J. & Jones, N. L. ( 2004;). Helicobacter pylori induces apoptosis of macrophages in association with alterations in the mitochondrial pathway. Infect Immun 72, 2889–2898.[CrossRef]
    [Google Scholar]
  40. Merrell, D. S. & Falkow, S. ( 2004;). Frontal and stealth attack strategies in microbial pathogenesis. Nature 430, 250–256.[CrossRef]
    [Google Scholar]
  41. Miehlke, S., Kirsch, C., Agha-Amiri, K., Gunther, T., Lehn, N., Malfertheiner, P., Stolte, M., Ehninger, G. & Bayerdorffer, E. ( 2000;). The Helicobacter pylori vacA s1, m1 genotype and cagA is associated with gastric carcinoma in Germany. Int J Cancer 87, 322–327.[CrossRef]
    [Google Scholar]
  42. Molinari, M., Galli, C., Norais, N., Telford, J., Rappuoli, R., Luzio, J. P. & Montecucco, C. ( 1997;). Vacuoles induced by Helicobacter pylori toxin contain both late endosomal and lysosomal markers. J Biol Chem 272, 25339–25344.[CrossRef]
    [Google Scholar]
  43. Molinari, M., Salio, M., Galli, C., Norais, N., Rappuoli, R., Lanzavecchia, A. & Montecucco, C. ( 1998a;). Selective inhibition of Ii-dependent antigen presentation by Helicobacter pylori toxin VacA. J Exp Med 187, 135–140.[CrossRef]
    [Google Scholar]
  44. Molinari, M., Galli, C., de Bernard, M., Norais, N., Ruysschaert, J.-M., Rappuoli, R. & Montecucco, C. ( 1998b;). The acid activation of Helicobacter pylori toxin VacA: structural and membrane binding studies. Biochem Biophys Res Comm 248, 334–340.[CrossRef]
    [Google Scholar]
  45. Moll, G., Papini, E., Colonna, R., Burroni, D., Telford, J., Rappuoli, R. & Montecucco, C. ( 1995;). Lipid interaction of the 37-kDa and 58-kDa fragments of the Helicobacter pylori cytotoxin. Eur J Biochem 234, 947–952.[CrossRef]
    [Google Scholar]
  46. Oomen, C. J., Van Ulsen, P., Van Gelder, P., Feijen, M., Tommassen, J. & Gros, P. ( 2004;). Structure of the translocator domain of a bacterial autotransporter. EMBO J 23, 1257–1266.[CrossRef]
    [Google Scholar]
  47. Pagliaccia, C., de Bernard, M., Lupetti, P. & 7 other authors ( 1998;). The m2 form of the Helicobacter pylori cytotoxin has cell type-specific vacuolating activity. Proc Natl Acad Sci U S A 95, 10212–10217.[CrossRef]
    [Google Scholar]
  48. Pagliaccia, C., Wang, X.-M., Tardy, F., Telford, J. L., Ruysschaert, J.-M. & Cabiaux, V. ( 2000;). Structure and interaction of VacA of Helicobacter pylori with a lipid membrane. Eur J Biochem 267, 104–109.[CrossRef]
    [Google Scholar]
  49. Papini, E., Satin, B., Norais, N., de Bernard, M., Telford, J. L., Rappuoli, R. & Montecucco, C. ( 1998;). Selective increase of the permeability of polarized epithelial cell monolayers by Helicobacter pylori vacuolating toxin. J Clin Invest 102, 813–820.[CrossRef]
    [Google Scholar]
  50. Patel, H. K., Willhite, D. C., Patel, R. M., Ye, D., Williams, C. L., Torres, E. M., Marty, K. B., MacDonald, R. A. & Blanke, S. R. ( 2002;). Plasma membrane cholesterol modulates cellular vacuolation induced by the Helicobacter pylori vacuolating cytotoxin. Infect Immun 70, 4112–4123.[CrossRef]
    [Google Scholar]
  51. Pelicic, V., Reyrat, J.-M., Sartori, L., Pagliaccia, C., Rappuoli, R., Telford, J. L., Montecucco, C. & Papini, E. ( 1999;). Helicobacter pylori VacA cytotoxin associated with the bacteria increases epithelial permeability independently of its vacuolating activity. Microbiology 145, 2043–2050.[CrossRef]
    [Google Scholar]
  52. Salama, N. R., Otto, G., Tompkins, L. & Falkow, S. ( 2001;). Vacuolating cytotoxin of Helicobacter pylori plays a role during colonization in a mouse model of infection. Infect Immun 69, 730–736.[CrossRef]
    [Google Scholar]
  53. Schmitt, W. & Haas, R. ( 1994;). Genetic analysis of the Helicobacter pylori vacuolating cytotoxin: structural similarities with the IgA protease type of exported protein. Mol Microbiol 12, 307–319.[CrossRef]
    [Google Scholar]
  54. Schraw, W., McClain, M. S. & Cover, T. L. ( 1999;). Kinetics and mechanisms of extracellular protein release by Helicobacter pylori. Infect Immun 67, 5247–5252.
    [Google Scholar]
  55. Schraw, W., Li, Y., McClain, M. S., van der Goot, F. G. & Cover, T. L. ( 2002;). Association of Helicobacter pylori vacuolating toxin (VacA) with lipid rafts. J Biol Chem 277, 34642–34650.[CrossRef]
    [Google Scholar]
  56. Seto, K., Hayashi-Kuwabara, Y., Yoneta, T., Suda, H. & Tamaki, H. ( 1998;). Vacuolation induced by cytotoxin from Helicobacter pylori is mediated by the EGF receptor in HeLa cells. FEBS Lett 431, 347–350.[CrossRef]
    [Google Scholar]
  57. Sijbrandi, R., Urbanus, M. L., ten Hagen-Jongman, C. M., Bernstein, H. D., Ougega, B., Otto, B. R. & Luirink, J. ( 2003;). Signal recognition particle (SRP)-mediated targeting and Sec-dependent translocation of an extracellular Escherichia coli protein. J Biol Chem 278, 4654–4659.[CrossRef]
    [Google Scholar]
  58. Sundrud, M. S., Torres, V. J., Unutmaz, D. & Cover, T. L. ( 2004;). Inhibition of primary human T cell proliferation by Helicobacter pylori vacuolating toxin (VacA) is independent of VacA effects on IL-2 secretion. Proc Natl Acad Sci U S A 101, 7727–7732.[CrossRef]
    [Google Scholar]
  59. Szabò, I., Brutsche, S., Tombola, F. & 7 other authors ( 1999;). Formation of anion-selective channels in the cell plasma membrane by the toxin VacA of Helicobacter pylori is required for its biological activity. EMBO J 18, 5517–5527.[CrossRef]
    [Google Scholar]
  60. Telford, J. L., Ghiara, P., Dell'Orco, M. & 7 other authors ( 1994;). Gene structure of the Helicobacter pylori cytotoxin and evidence of its key role in gastric disease. J Exp Med 179, 1653–1658.[CrossRef]
    [Google Scholar]
  61. Tombola, F., Carlesso, C., Szabò, I. & 7 other authors ( 1999;). Helicobacter pylori vacuolating cytotoxin forms anion-selective channels in planar lipid bilayers: possible implications for the mechanism of cellular vacuolation. Biophys J 76, 1401–1409.[CrossRef]
    [Google Scholar]
  62. Van Doorn, L.-J., Figueiredo, C., Sanna, R., Pena, S., Midolo, P., Ng, E. K. W., Atherton, J. C., Blaser, M. J. & Quint, W. G. V. ( 1998;). Expanding allelic diversity of Helicobacter pylori vacA. J Clin Microbiol 36, 2597–2603.
    [Google Scholar]
  63. Veiga, E., Sugawara, E., Nikaido, H., de Lorenzo, V. & Fernandez, L. A. ( 2002;). Export of autotransported proteins proceeds through an oligomeric ring shaped by C-terminal domains. EMBO J 21, 2122–2131.[CrossRef]
    [Google Scholar]
  64. Vinion-Dubiel, A. D., McClain, M. S., Czajkowsky, D. M. & 8 other authors ( 1999;). A dominant negative mutant of Helicobacter pylori vacuolating toxin (VacA) inhibits VacA-induced cell vacuolation. J Biol Chem 274, 37736–37742.[CrossRef]
    [Google Scholar]
  65. Voulhoux, R., Bos, M. P., Geurtsen, J., Mols, M. & Tommassen, J. ( 2003;). Role of a highly conserved bacterial protein in outer membrane protein assembly. Science 299, 262–265.[CrossRef]
    [Google Scholar]
  66. Willhite, D. C. & Blanke, S. R. ( 2004;). Helicobacter pylori vacuolating cytotoxin enters cells, localizes to the mitochondria, and induces mitochondrial membrane permeability changes correlated to toxin channel activity. Cell Microbiol 6, 143–154.[CrossRef]
    [Google Scholar]
  67. Willhite, D. C., Cover, T. L. & Blanke, S. R. ( 2003;). Cellular vacuolation and mitochondrial cytochrome c release are independent outcomes of Helicobacter pylori vacuolating cytotoxin activity that are each dependent on membrane channel formation. J Biol Chem 278, 48204–48209.[CrossRef]
    [Google Scholar]
  68. Wood, W. B. ( 1966;). Host specificity of DNA produced by Escherichia coli: bacterial mutation affecting the restriction and modification of DNA. J Mol Biol 16, 118–133.[CrossRef]
    [Google Scholar]
  69. Yahiro, K., Niidome, T., Kimura, M. & 7 other authors ( 1999;). Activation of Helicobacter pylori VacA toxin by alkaline or acid conditions increases its binding to a 250-kDa receptor protein-tyrosine phosphatase β. J Biol Chem 274, 36693–36699.[CrossRef]
    [Google Scholar]
  70. Yahiro, K., Wada, A., Nakayama, M. & 8 other authors ( 2003;). Protein-tyrosine phosphatase α, RPTP α, is a Helicobacter pylori VacA receptor. J Biol Chem 278, 19183–19189.[CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.45946-0
Loading
/content/journal/jmm/10.1099/jmm.0.45946-0
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