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Volume 156, Issue 8

Efa-1/LifA mediates intestinal colonization of calves by enterohaemorrhagic O26 : H– in a manner independent of glycosyltransferase and cysteine protease motifs or effects on type III secretion Free

Abstract

Enterohaemorrhagic (EHEC) comprise a group of animal and zoonotic pathogens of worldwide importance. Our previous research established that intestinal colonization of calves by EHEC serotypes O5 : H– and O111 : H– requires HEC actor for dherence (Efa-1), also known as lymphostatin (LifA). Towards an understanding of the mode of action of Efa-1/LifA, chromosomal in-frame deletions of predicted glycosyltransferase (DXD) and cysteine protease (CHD) motifs were created in a Δ derivative of EHEC O26 : H–. The magnitude and duration of faecal excretion of EHEC O26 : H– were significantly reduced by null mutation of /, but were not impaired by ΔDXD or ΔCHD mutations, in contrast to observations made with truncated Efa-1/LifA mutants of in mice. Although Efa-1/LifA influences the induction of colonic hyperplasia in mice, EHEC O26 : H– Efa-1/LifA was not required for fluid accumulation or neutrophil recruitment in bovine ileal loops. In contrast to observations with EHEC O5 : H– or O111 : H– mutants, inactivation of / in EHEC O26 : H– did not significantly affect adherence or secretion of type III secreted proteins that play pivotal roles in calf colonization. Lymphostatin activity could not be reliably demonstrated in lysates of EHEC O26 : H–; however, deletion of the glycosyltransferase and cysteine protease motifs in Efa-1/LifA from enteropathogenic O127 : H6 abolished lymphostatin activity. Our data uncouple the role of Efa-1/LifA in calf colonization from effects on type III secretion and reinforce the potential for pathotype- and serotype-specific phenotypes.

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Keyword(s): EHEC, enterohaemorrhagic E. coli , EPEC, enteropathogenic E. coli , LEE, locus of enterocyte effacement , PBL, peripheral blood lymphocyte , PMN, polymorphonuclear and T3SS, type III secretion system
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2010-08-01
2024-04-18
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References

  1. Abu-Median A. B., van Diemen P. M., Dziva F., Vlisidou I., Wallis T. S., Stevens M. P. 2006; Functional analysis of lymphostatin homologues in enterohaemorrhagic Escherichia coli. FEMS Microbiol Lett 258:43–49
     [Google Scholar]
  2. Afset J. E., Bruant G., Brousseau R., Harel J., Anderssen E., Bevanger L., Bergh K. 2006; Identification of virulence genes linked with diarrhea due to atypical enteropathogenic Escherichia coli by DNA microarray analysis and PCR. J Clin Microbiol 44:3703–3711
     [Google Scholar]
  3. Arbeloa A., Bulgin R. R., MacKenzie G., Shaw R. K., Pallen M. J., Crepin V. F., Berger C. N., Frankel G. 2008; Subversion of actin dynamics by EspM effectors of attaching and effacing bacterial pathogens. Cell Microbiol 10:1429–1441
     [Google Scholar]
  4. Arbeloa A., Garnett J., Lillington J., Bulgin R. R., Berger C. N., Lea S. M., Matthews S., Frankel G. 2010; EspM2 is a RhoA guanine nucleotide exchange factor. Cell Microbiol 12:654–664
     [Google Scholar]
  5. Babbin B. A., Sasaki M., Gerner-Schmidt K. W., Nusrat A., Klapproth J. M. 2009; The bacterial virulence factor lymphostatin compromises intestinal epithelial barrier function by modulating rho GTPases. Am J Pathol 174:1347–1357
     [Google Scholar]
  6. Badea L., Doughty S., Nicholls L., Sloan J., Robins-Browne R. M., Hartland E. L. 2003; Contribution of Efa1/LifA to the adherence of enteropathogenic Escherichia coli to epithelial cells. Microb Pathog 34:205–215
     [Google Scholar]
  7. Belland R. J., Scidmore M. A., Crane D. D., Hogan D. M., Whitmire W., McClarty G., Caldwell H. D. 2001; Chlamydia trachomatis cytotoxicity associated with complete and partial cytotoxin genes. Proc Natl Acad Sci U S A 98:13984–13989
     [Google Scholar]
  8. Bettelheim K. A. 2007; The non-O157 shiga-toxigenic (verocytotoxigenic) Escherichia coli; under-rated pathogens. Crit Rev Microbiol 33:67–87
     [Google Scholar]
  9. Bielaszewska M., Sonntag A. K., Schmidt M. A., Karch H. 2007; Presence of virulence and fitness gene modules of enterohemorrhagic Escherichia coli in atypical enteropathogenic Escherichia coli O26. Microbes Infect 9:891–897
     [Google Scholar]
  10. Busch C., Hofmann F., Selzer J., Munro S., Jeckel D., Aktories K. 1998; A common motif of eukaryotic glycosyltransferases is essential for the enzyme activity of large clostridial cytotoxins. J Biol Chem 273:19566–19572
     [Google Scholar]
  11. Cleary J., Lai L. C., Shaw R. K., Straatman-Iwanowska A., Donnenberg M. S., Frankel G., Knutton S. 2004; Enteropathogenic Escherichia coli (EPEC) adhesion to intestinal epithelial cells: role of bundle-forming pili (BFP), EspA filaments and intimin. Microbiology 150:527–538
     [Google Scholar]
  12. 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]
  13. Dean-Nystrom E. A., Bosworth B. T., Moon H. W., O'Brien A. D. 1998; Escherichia coli O157 : H7 requires intimin for enteropathogenicity in calves. Infect Immun 66:4560–4563
     [Google Scholar]
  14. Dziva F., van Diemen P. M., Stevens M. P., Smith A. J., Wallis T. S. 2004; Identification of Escherichia coli O157 : H7 genes influencing colonization of the bovine gastrointestinal tract using signature-tagged mutagenesis. Microbiology 150:3631–3645
     [Google Scholar]
  15. Dziva F., Mahajan A., Cameron P., Currie C., McKendrick I. J., Wallis T. S., Smith D. G., Stevens M. P. 2007; EspP, a Type V-secreted serine protease of enterohaemorrhagic Escherichia coli O157 : H7, influences intestinal colonization of calves and adherence to bovine primary intestinal epithelial cells. FEMS Microbiol Lett 271:258–264
     [Google Scholar]
  16. Ebel F., Podzadel T., Rohde M., Kresse A. U., Krämer S., Deibel C., Guzmán 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]
  17. Gyles C. L. 2007; Shiga toxin-producing Escherichia coli: an overview. J Anim Sci 85:E45–E62
     [Google Scholar]
  18. Hofmann F., Busch C., Prepens U., Just I., Aktories K. 1997; Localization of the glucosyltransferase activity of Clostridium difficile toxin B to the N-terminal part of the holotoxin. J Biol Chem 272:11074–11078
     [Google Scholar]
  19. Horton R. M., Hunt H. D., Ho S. N., Pullen J. K., Pease L. R. 1989; Engineering hybrid genes without the use of restriction enzymes: gene splicing by overlap extension. Gene 77:61–68
     [Google Scholar]
  20. Karmali M. A., Mascarenhas M., Shen S., Ziebell K., Johnson S., Reid-Smith R., Isaac-Renton J., Clark C., Rahn K., Kaper J. B. 2003; Association of genomic O island 122 of Escherichia coli EDL933 with verocytotoxin-producing Escherichia coli seropathotypes that are linked to epidemic and/or serious disease. J Clin Microbiol 41:4930–4940
     [Google Scholar]
  21. Klapproth J. M., Donnenberg M. S., Abraham J. M., Mobley H. L., James S. P. 1995; Products of enteropathogenic Escherichia coli inhibit lymphocyte activation and lymphokine production. Infect Immun 63:2248–2254
     [Google Scholar]
  22. Klapproth J. M., Donnenberg M. S., Abraham J. M., James S. P. 1996; Products of enteropathogenic E. coli inhibit lymphokine production by gastrointestinal lymphocytes. Am J Physiol 271:G841–G848
     [Google Scholar]
  23. Klapproth J. M., Scaletsky I. C., McNamara B. P., Lai L. C., Malstrom C., James S. P., Donnenberg M. S. 2000; A large toxin from pathogenic Escherichia coli strains that inhibits lymphocyte activation. Infect Immun 68:2148–2155
     [Google Scholar]
  24. Klapproth J. M., Sasaki M., Sherman M., Babbin B., Donnenberg M. S., Fernandes P. J., Scaletsky I. C., Kalman D., Nusrat A., Williams I. R. 2005; Citrobacter rodentium lifA/ efa1 is essential for colonic colonization and crypt cell hyperplasia in vivo. Infect Immun 73:1441–1451
     [Google Scholar]
  25. Levine M. M., Berquist E. J., Nalin D. R., Waterman D. H., Hornick R. B., Young C. R., Stoman S. 1978; Escherichia coli that cause diarrhoea but do not produce heat-labile or heat-stable enterotoxins and are non-invasive. Lancet 1:1119–1122
     [Google Scholar]
  26. Low A. S., Dziva F., Torres A. G., Martinez J. L., Rosser T., Naylor S., Spears K., Holden N., Mahajan A. other authors 2006; Cloning, expression, and characterization of fimbrial operon F9 from enterohemorrhagic Escherichia coli O157 : H7. Infect Immun 74:2233–2244
     [Google Scholar]
  27. Malstrom C., James S. 1998; Inhibition of murine splenic and mucosal lymphocyte function by enteric bacterial products. Infect Immun 66:3120–3127
     [Google Scholar]
  28. Menge C., Wieler L. H., Schlapp T., Baljer G. 1999; Shiga toxin 1 from Escherichia coli blocks activation and proliferation of bovine lymphocyte subpopulations in vitro. Infect Immun 67:2209–2217
     [Google Scholar]
  29. Menge C., Stamm I., van Diemen P. M., Sopp P., Baljer G., Wallis T. S., Stevens M. P. 2004; Phenotypic and functional characterization of intraepithelial lymphocytes in a bovine ligated intestinal loop model of enterohaemorrhagic Escherichia coli infection. J Med Microbiol 53:573–579
     [Google Scholar]
  30. Milton D. L., O'Toole R., Horstedt P., Wolf-Watz H. 1996; Flagellin A is essential for the virulence of Vibrio anguillarum. J Bacteriol 178:1310–1319
     [Google Scholar]
  31. Morabito S., Tozzoli R., Oswald E., Caprioli A. 2003; A mosaic pathogenicity island made up of the locus of enterocyte effacement and a pathogenicity island of Escherichia coli O157 : H7 is frequently present in attaching and effacing E. coli. Infect Immun 71:3343–3348
     [Google Scholar]
  32. Naylor S. W., Roe A. J., Nart P., Spears K., Smith D. G., Low J. C., Gally D. L. 2005; Escherichia coli O157 : H7 forms attaching and effacing lesions at the terminal rectum of cattle and colonization requires the LEE4 operon. Microbiology 151:2773–2781
     [Google Scholar]
  33. Nicholls L., Grant T. H., Robins-Browne R. M. 2000; Identification of a novel genetic locus that is required for in vitro adhesion of a clinical isolate of enterohaemorrhagic Escherichia coli to epithelial cells. Mol Microbiol 35:275–288
     [Google Scholar]
  34. Ogura Y., Ooka T., Iguchi A., Toh H., Asadulghani M., Oshima K., Kodama T., Abe H., Nakayama K. other authors 2009; Comparative genomics reveal the mechanism of the parallel evolution of O157 and non-O157 enterohemorrhagic Escherichia coli. Proc Natl Acad Sci U S A 106:17939–17944
     [Google Scholar]
  35. Robinson C. M., Sinclair J. F., Smith M. J., O'Brien A. D. 2006; Shiga toxin of enterohemorrhagic Escherichia coli type O157 : H7 promotes intestinal colonization. Proc Natl Acad Sci U S A 103:9667–9672
     [Google Scholar]
  36. Shao F., Merritt P. M., Bao Z., Innes R. W., Dixon J. E. 2002; A Yersinia effector and a Pseudomonas avirulence protein define a family of cysteine proteases functioning in bacterial pathogenesis. Cell 109:575–588
     [Google Scholar]
  37. Simon R., Preifer U., Puhler A. 1983; A broad host range mobilisation system for in vivo genetic engineering: transposon mutagenesis in Gram-negative bacteria. Biotechnology (N Y) 1:784–791
     [Google Scholar]
  38. Stevens M. P., Marchès O., Campbell J., Huter V., Frankel G., Phillips A. D., Oswald E., Wallis T. S. 2002a; Intimin, Tir, and Shiga toxin 1 do not influence enteropathogenic responses to Shiga toxin-producing Escherichia coli in bovine ligated intestinal loops. Infect Immun 70:945–952
     [Google Scholar]
  39. Stevens M. P., van Diemen P. M., Frankel G., Phillips A. D., Wallis T. S. 2002b; Efa1 influences colonization of the bovine intestine by Shiga toxin-producing Escherichia coli serotypes O5 and O111. Infect Immun 70:5158–5166
     [Google Scholar]
  40. Stevens M. P., Roe A. J., Vlisidou I., van Diemen P. M., La Ragione R. M., Best A., Woodward M. J., Gally D. L., Wallis T. S. 2004; Mutation of toxB and a truncated version of the efa-1 gene in Escherichia coli O157 : H7 influences the expression and secretion of locus of enterocyte effacement-encoded proteins but not intestinal colonization in calves or sheep. Infect Immun 72:5402–5411
     [Google Scholar]
  41. Tatsuno I., Horie M., Abe H., Miki T., Makino K., Shinagawa H., Taguchi H., Kamiya S., Hayashi T., Sasakawa C. 2001; toxB gene on pO157 of enterohemorrhagic Escherichia coli O157 : H7 is required for full epithelial cell adherence phenotype. Infect Immun 69:6660–6669
     [Google Scholar]
  42. Tozzoli R., Caprioli A., Morabito S. 2005; Detection of toxB, a plasmid virulence gene of Escherichia coli O157, in enterohemorrhagic and enteropathogenic E. coli. J Clin Microbiol 43:4052–4056
     [Google Scholar]
  43. van Diemen P. M., Dziva F., Stevens M. P., Wallis T. S. 2005; Identification of enterohemorrhagic Escherichia coli O26 : H– genes required for intestinal colonization in calves. Infect Immun 73:1735–1743
     [Google Scholar]
  44. van Diemen P. M., Dziva F., Abu-Median A., Wallis T. S., van den Bosch H., Dougan G., Chanter N., Frankel G., Stevens M. P. 2007; Subunit vaccines based on intimin and Efa-1 polypeptides induce humoral immunity in cattle but do not protect against intestinal colonisation by enterohaemorrhagic Escherichia coli O157 : H7 or O26 : H–. Vet Immunol Immunopathol 116:47–58
     [Google Scholar]
  45. Vlisidou I., Lyte M., van Diemen P. M., Hawes P., Monaghan P., Wallis T. S., Stevens M. P. 2004; The neuroendocrine stress hormone norepinephrine augments Escherichia coli O157 : H7-induced enteritis and adherence in a bovine ligated ileal loop model of infection. Infect Immun 72:5446–5451
     [Google Scholar]
  46. Vlisidou I., Dziva F., La Ragione R. M., Best A., Garmendia J., Hawes P., Monaghan P., Cawthraw S. A., Frankel G. other authors 2006; Role of intimin-Tir interactions and the Tir-cytoskeleton coupling protein in the colonization of calves and lambs by Escherichia coli O157 : H7. Infect Immun 74:758–764
     [Google Scholar]
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Efa-1/LifA mediates intestinal colonization of calves by enterohaemorrhagic Escherichia coli O26 : H– in a manner independent of glycosyltransferase and cysteine protease motifs or effects on type III secretion
Microbiology 156, 2527 (2010); https://doi.org/10.1099/mic.0.039685-0
/content/journal/micro/10.1099/mic.0.039685-0
/content/journal/micro/10.1099/mic.0.039685-0
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