1887

Abstract

is a leading cause of nosocomial infections, causing a spectrum of diseases ranging from diarrhoea to pseudomembranous colitis triggered by a range of virulence factors including toxins A (TcdA) and B (TcdB). TcdA and TcdB are monoglucosyltransferases that irreversibly glycosylate small Rho GTPases, inhibiting their ability to interact with their effectors, guanine nucleotide exchange factors, and membrane partners, leading to disruption of downstream signalling pathways and cell death. In addition, TcdB targets the mitochondria, inducing the intrinsic apoptotic pathway resulting in TcdB-mediated apoptosis. Modulation of apoptosis is a common strategy used by infectious agents. Recently, we have shown that the enteropathogenic (EPEC) type III secretion system effector NleH has a broad-range anti-apoptotic activity. In this study we examined the effects of NleH on cells challenged with TcdB. During infection with wild-type EPEC, NleH inhibited TcdB-induced apoptosis at both low and high toxin concentrations. Transfected alone was sufficient to block TcdB-induced cell rounding, nuclear condensation, mitochondrial swelling and lysis, and activation of caspase-3. These results show that NleH acts via a global anti-apoptotic pathway.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.037259-0
2010-06-01
2024-04-19
Loading full text...

Full text loading...

/deliver/fulltext/micro/156/6/1815.html?itemId=/content/journal/micro/10.1099/mic.0.037259-0&mimeType=html&fmt=ahah

References

  1. Bartlett J. G., Moon N., Chang T. W., Taylor N., Onderdonk A. B. 1978; Role of Clostridium difficile in antibiotic-associated Pseudomembranous colitis. Gastroenterology 75:778–782
    [Google Scholar]
  2. Budihardjo I., Oliver H., Lutter M., Luo X., Wang X. 1999; Biochemical pathways of caspase activation during apoptosis. Annu Rev Cell Dev Biol 15:269–290
    [Google Scholar]
  3. Caron E., Crepin V. F., Simpson N., Knutton S., Garmendia J., Frankel G. 2006; Subversion of actin dynamics by EPEC and EHEC. Curr Opin Microbiol 9:40–45
    [Google Scholar]
  4. Chae H. J., Kim H. R., Xu C., Bailly-Maitre B., Krajewska M., Krajewski S., Banares S., Cui J., Digicaylioglu M. other authors 2004; BI-1 regulates an apoptosis pathway linked to endoplasmic reticulum stress. Mol Cell 15:355–366
    [Google Scholar]
  5. Chen H. D., Frankel G. 2005; Enteropathogenic Escherichia coli: unravelling pathogenesis. FEMS Microbiol Rev 29:83–98
    [Google Scholar]
  6. Crane J. K., Majumdar S., Pickhardt D. F. III 1999; Host cell death due to enteropathogenic Escherichia coli has features of apoptosis. Infect Immun 67:2575–2584
    [Google Scholar]
  7. Crane J. K., McNamara B. P., Donnenberg M. S. 2001; Role of EspF in host cell death induced by enteropathogenic Escherichia coli. Cell Microbiol 3:197–211
    [Google Scholar]
  8. Desagher S., Martinou J. C. 2000; Mitochondria as the central control point of apoptosis. Trends Cell Biol 10:369–377
    [Google Scholar]
  9. 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]
  10. Gao X., Wan F., Mateo K., Callegari E., Wang D., Deng W., Puente J., Li F., Chaussee M. S. other authors 2009; Bacterial effector binding to ribosomal protein s3 subverts NF-kappaB function. PLoS Pathog 5:e1000708
    [Google Scholar]
  11. Garmendia J., Phillips A. D., Carlier M. F., Chong Y., Schüller S., Marches O., Dahan S., Oswald E., Shaw R. K. other authors 2004; TccP is an enterohaemorrhagic Escherichia coli O157 : H7 type III effector protein that couples Tir to the actin-cytoskeleton. Cell Microbiol 6:1167–1183
    [Google Scholar]
  12. Garmendia J., Frankel G., Crepin V. F. 2005; Enteropathogenic and enterohemorrhagic Escherichia coli infections: translocation, translocation, translocation. Infect Immun 73:2573–2585
    [Google Scholar]
  13. Genth H., Aktories K., Just I. 1999; Monoglucosylation of RhoA at threonine 37 blocks cytosol-membrane cycling. J Biol Chem 274:29050–29056
    [Google Scholar]
  14. Genth H., Dreger S. C., Huelsenbeck J., Just I. 2008; Clostridium difficile toxins: more than mere inhibitors of Rho proteins. Int J Biochem Cell Biol 40:592–597
    [Google Scholar]
  15. George W. L., Sutter V. L., Goldstein E. J., Ludwig S. L., Finegold S. M. 1978; Aetiology of antimicrobial-agent-associated colitis. Lancet 1:802–803
    [Google Scholar]
  16. Gross A., Pilcher K., Blachly-Dyson E., Basso E., Jockel J., Bassik M. C., Korsmeyer S. J., Forte M. 2000; Biochemical and genetic analysis of the mitochondrial response of yeast to BAX and BCL-X(L. Mol Cell Biol 20:3125–3136
    [Google Scholar]
  17. Hemrajani C., Berger C. N., Robinson K. S., Marches O., Mousnier A., Frankel G. 2010; NleH effectors interact with Bax inhibitor-1 to block apoptosis during enteropathogenic Escherichia coli infection. Proc Natl Acad Sci U S A 107:3129–3134
    [Google Scholar]
  18. Herrmann C., Ahmadian M. R., Hofmann F., Just I. 1998; Functional consequences of monoglucosylation of Ha-Ras at effector domain amino acid threonine 35. J Biol Chem 273:16134–16139
    [Google Scholar]
  19. Iguchi A., Thomson N. R., Ogura Y., Saunders D., Ooka T., Henderson I. R., Harris D., Asadulghani M., Kurokawa K. other authors 2009; Complete genome sequence and comparative genome analysis of enteropathogenic Escherichia coli O127 : H6 strain E2348/69. J Bacteriol 191:347–354
    [Google Scholar]
  20. Just I., Gerhard R. 2004; Large clostridial cytotoxins. Rev Physiol Biochem Pharmacol 152:23–47
    [Google Scholar]
  21. Just I., Selzer J., Wilm M., von Eichel-Streiber C., Mann M., Aktories K. 1995; Glucosylation of Rho proteins by Clostridium difficile toxin B. Nature 375:500–503
    [Google Scholar]
  22. Kato H., Kato N., Watanabe K., Iwai N., Nakamura H., Yamamoto T., Suzuki K., Kim S. M., Chong Y., Wasito E. B. 1998; Identification of toxin A-negative, toxin B-positive Clostridium difficile by PCR. J Clin Microbiol 36:2178–2182
    [Google Scholar]
  23. 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]
  24. Kroemer G., Reed J. C. 2000; Mitochondrial control of cell death. Nat Med 6:513–519
    [Google Scholar]
  25. Kuijper E. J., de Weerdt J., Kato H., Kato N., van Dam A. P., van der Vorm E. R., Weel J., van Rheenen C., Dankert J. 2001; Nosocomial outbreak of Clostridium difficile-associated diarrhoea due to a clindamycin-resistant enterotoxin A-negative strain. Eur J Clin Microbiol Infect Dis 20:528–534
    [Google Scholar]
  26. Kyne L., Hamel M. B., Polavaram R., Kelly C. P. 2002; Health care costs and mortality associated with nosocomial diarrhea due to Clostridium difficile. Clin Infect Dis 34:346–353
    [Google Scholar]
  27. Le S. S., Loucks F. A., Udo H., Richardson-Burns S., Phelps R. A., Bouchard R. J., Barth H., Aktories K., Tyler K. L. other authors 2005; Inhibition of Rac GTPase triggers a c-Jun- and Bim-dependent mitochondrial apoptotic cascade in cerebellar granule neurons. J Neurochem 94:1025–1039
    [Google Scholar]
  28. Levine M. M., Berquist E. J., Nalin D. R., Waterman D. H., Hornick R. B., Young C. R., Stoman S., Rowe B. 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]
  29. Limaye A. P., Turgeon D. K., Cookson B. T., Fritsche T. R. 2000; Pseudomembranous colitis caused by a toxin A(−) B(+) strain of Clostridium difficile. J Clin Microbiol 38:1696–1697
    [Google Scholar]
  30. Lyras D., O'Connor J. R., Howarth P. M., Sambol S. P., Carter G. P., Phumoonna T., Poon R., Adams V., Vedantam G. other authors 2009; Toxin B is essential for virulence of Clostridium difficile. Nature 458:1176–1179
    [Google Scholar]
  31. Mani N., Lyras D., Barroso L., Howarth P., Wilkins T., Rood J. I., Sonenshein A. L., Dupuy B. 2002; Environmental response and autoregulation of Clostridium difficile TxeR, a sigma factor for toxin gene expression. J Bacteriol 184:5971–5978
    [Google Scholar]
  32. Matamouros S., England P., Dupuy B. 2007; Clostridium difficile toxin expression is inhibited by the novel regulator TcdC. Mol Microbiol 64:1274–1288
    [Google Scholar]
  33. Matarrese P., Gambardella L., Cassone A., Vella S., Cauda R., Malorni W. 2003; Mitochondrial membrane hyperpolarization hijacks activated T lymphocytes toward the apoptotic-prone phenotype: homeostatic mechanisms of HIV protease inhibitors. J Immunol 170:6006–6015
    [Google Scholar]
  34. Matarrese P., Falzano L., Fabbri A., Gambardella L., Frank C., Geny B., Popoff M. R., Malorni W., Fiorentini C. 2007; Clostridium difficile toxin B causes apoptosis in epithelial cells by thrilling mitochondria. Involvement of ATP-sensitive mitochondrial potassium channels. J Biol Chem 282:9029–9041
    [Google Scholar]
  35. 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
    [Google Scholar]
  36. Nataro J. P., Kaper J. B. 1998; Diarrheagenic Escherichia coli. Clin Microbiol Rev 11:142–201
    [Google Scholar]
  37. Nougayrede J. P., Foster G. H., Donnenberg M. S. 2007; Enteropathogenic Escherichia coli effector EspF interacts with host protein Abcf2. Cell Microbiol 9:680–693
    [Google Scholar]
  38. Reineke J., Tenzer S., Rupnik M., Koschinski A., Hasselmayer O., Schrattenholz A., Schild H., von Eichel-Streiber C. 2007; Autocatalytic cleavage of Clostridium difficile toxin B. Nature 446:415–419
    [Google Scholar]
  39. Rupnik M., Wilcox M. H., Gerding D. N. 2009; Clostridium difficile infection: new developments in epidemiology and pathogenesis. Nat Rev Microbiol 7:526–536
    [Google Scholar]
  40. 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]
  41. Sehr P., Joseph G., Genth H., Just I., Pick E., Aktories K. 1998; Glucosylation and ADP ribosylation of rho proteins: effects on nucleotide binding, GTPase activity, and effector coupling. Biochemistry 37:5296–5304
    [Google Scholar]
  42. Tobe T., Beatson S. A., Taniguchi H., Abe H., Bailey C. M., Fivian A., Younis R., Matthews S., Marches O. other authors 2006; An extensive repertoire of type III secretion effectors in Escherichia coli O157 and the role of lambdoid phages in their dissemination. Proc Natl Acad Sci U S A 103:14941–14946
    [Google Scholar]
  43. Voth D. E., Ballard J. D. 2005; Clostridium difficile toxins: mechanism of action and role in disease. Clin Microbiol Rev 18:247–263
    [Google Scholar]
  44. Xu Q., Reed J. C. 1998; Bax inhibitor-1, a mammalian apoptosis suppressor identified by functional screening in yeast. Mol Cell 1:337–346
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.037259-0
Loading
/content/journal/micro/10.1099/mic.0.037259-0
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