This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Ching, J. C. Y.
Right arrow Articles by Sherman, P. M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Ching, J. C. Y.
Right arrow Articles by Sherman, P. M.

 Previous Article  |  Next Article 

Infection and Immunity, August 2002, p. 4669-4677, Vol. 70, No. 8
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.8.4669-4677.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Escherichia coli Shiga-Like Toxins Induce Apoptosis and Cleavage of Poly(ADP-Ribose) Polymerase via In Vitro Activation of Caspases

Joyce C. Y. Ching,1,2 Nicola L. Jones,1,3,4 Peter J. M. Ceponis,1,5 Mohamed A. Karmali,6 and Philip M. Sherman1,2,3,5*

Research Institute, Hospital for Sick Children,1 Institute of Medical Science,2 Departments of Pediatrics,3 Physiology,4 Laboratory Medicine & Pathobiology, University of Toronto, Toronto,5 Health Canada, Guelph, Ontario, Canada6

Received 15 January 2002/ Returned for modification 28 February 2002/ Accepted 23 April 2002

Shiga-like toxin-producing Escherichia coli causes hemorrhagic colitis and hemolytic-uremic syndrome in association with the production of Shiga-like toxins, which induce cell death via either necrosis or apoptosis. However, the abilities of different Shiga-like toxins to trigger apoptosis and the sequence of intracellular signaling events mediating the death of epithelial cells have not been completely defined. Fluorescent dye staining with acridine orange and ethidium bromide showed that Shiga-like toxin 1 (Stx1) induced apoptosis of HEp-2 cells in a dose- and time-dependent manner. Stx2 also induced apoptosis in a dose-dependent manner. Apoptosis induced by Stx1 (200 ng/ml) and apoptosis induced by Stx2 (200 ng/ml) were maximal following incubation with cells for 24 h (94.3% ± 1.8% and 81.7% ± 5.2% of the cells, respectively). Toxin-treated cells showed characteristic features of apoptosis, including membrane blebbing, DNA fragmentation, chromatin condensation, cell shrinkage, and the formation of apoptotic bodies, as assessed by transmission electron microscopy. Stx2c induced apoptosis weakly even at a high dose (1,000 ng/ml for 24 h; 26.7% ± 1.3% of the cells), whereas Stx2e did not induce apoptosis of HEp-2 cells. Thin-layer chromatography confirmed that HEp-2 cells express the Stx1-Stx2-Stx2c receptor, globotriaosylceramide (Gb3), but not the Stx2e receptor, globotetraosylceramide (Gb4). Western blot analysis of poly(ADP-ribose) polymerase (PARP), a DNA repair enzyme, demonstrated that incubation with Stx1 and Stx2 induced cleavage, whereas incubation with Stx2e did not result in cleavage of PARP. A pan-caspase inhibitor (Z-VAD-FMK) and a caspase-8-specific inhibitor (Z-IETD-FMK) eliminated, in a dose-dependent fashion, the cleavage of PARP induced by Shiga-like toxins. Caspase-8 activation was confirmed by detection of cleavage of this enzyme by immunoblotting. Cleavage of caspase-9 and the proapoptotic member of the Bcl-2 family BID was also induced by Stx1, as determined by immunoblot analyses. We conclude that different Shiga-like toxins induce different degrees of apoptosis that correlates with toxin binding to the glycolipid receptor Gb3 and that caspases play an integral role in the signal transduction cascade leading to toxin-mediated programmed cell death.

* Corresponding author. Mailing address: Rm. 8411, Hospital for Sick Children, 555 University Ave., Toronto, Ontario M5G 1X8, Canada. Phone: (416) 813-7734. Fax: (416) 813-6531. E-mail: sherman{at}sickkids.ca.

Editor: J. T. Barbieri

Infection and Immunity, August 2002, p. 4669-4677, Vol. 70, No. 8
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.8.4669-4677.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • 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. USA 103: 9667-9672 [Abstract] [Full Text]  
  • Nazli, A., Wang, A., Steen, O., Prescott, D., Lu, J., Perdue, M. H., Soderholm, J. D., Sherman, P. M., McKay, D. M. (2006). Enterocyte Cytoskeleton Changes Are Crucial for Enhanced Translocation of Nonpathogenic Escherichia coli across Metabolically Stressed Gut Epithelia. Infect. Immun. 74: 192-201 [Abstract] [Full Text]  
  • Nakayama, K., Pergolizzi, R. G., Crystal, R. G. (2005). Gene Transfer-Mediated Pre-mRNA Segmental Trans-splicing As a Strategy to Deliver Intracellular Toxins for Cancer Therapy. Cancer Res. 65: 254-263 [Abstract] [Full Text]  
  • Sukumaran, S. K., Selvaraj, S. K., Prasadarao, N. V. (2004). Inhibition of Apoptosis by Escherichia coli K1 Is Accompanied by Increased Expression of BclXL and Blockade of Mitochondrial Cytochrome c Release in Macrophages. Infect. Immun. 72: 6012-6022 [Abstract] [Full Text]  
  • Menaker, R. J., Ceponis, P. J. M., Jones, N. L. (2004). Helicobacter pylori Induces Apoptosis of Macrophages in Association with Alterations in the Mitochondrial Pathway. Infect. Immun. 72: 2889-2898 [Abstract] [Full Text]  
  • Ohara, M., Hayashi, T., Kusunoki, Y., Miyauchi, M., Takata, T., Sugai, M. (2004). Caspase-2 and Caspase-7 Are Involved in Cytolethal Distending Toxin-Induced Apoptosis in Jurkat and MOLT-4 T-Cell Lines. Infect. Immun. 72: 871-879 [Abstract] [Full Text]  
  • Mitchell, D. J., Huynh, H. Q., Ceponis, P. J. M., Jones, N. L., Sherman, P. M. (2004). Helicobacter pylori Disrupts STAT1-Mediated Gamma Interferon-Induced Signal Transduction in Epithelial Cells. Infect. Immun. 72: 537-545 [Abstract] [Full Text]  
  • Tetaud, C., Falguieres, T., Carlier, K., Lecluse, Y., Garibal, J., Coulaud, D., Busson, P., Steffensen, R., Clausen, H., Johannes, L., Wiels, J. (2003). Two Distinct Gb3/CD77 Signaling Pathways Leading to Apoptosis Are Triggered by Anti-Gb3/CD77 mAb and Verotoxin-1. J. Biol. Chem. 278: 45200-45208 [Abstract] [Full Text]  
  • Ceponis, P. J. M., McKay, D. M., Menaker, R. J., Galindo-Mata, E., Jones, N. L. (2003). Helicobacter pylori Infection Interferes with Epithelial Stat6-Mediated Interleukin-4 Signal Transduction Independent of cagA, cagE, or VacA. J. Immunol. 171: 2035-2041 [Abstract] [Full Text]  
  • Ceponis, P. J. M., McKay, D. M., Ching, J. C. Y., Pereira, P., Sherman, P. M. (2003). Enterohemorrhagic Escherichia coli O157:H7 Disrupts Stat1-Mediated Gamma Interferon Signal Transduction in Epithelial Cells. Infect. Immun. 71: 1396-1404 [Abstract] [Full Text]  
  • Smith, W. E., Kane, A. V., Campbell, S. T., Acheson, D. W. K., Cochran, B. H., Thorpe, C. M. (2003). Shiga Toxin 1 Triggers a Ribotoxic Stress Response Leading to p38 and JNK Activation and Induction of Apoptosis in Intestinal Epithelial Cells. Infect. Immun. 71: 1497-1504 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»