Document Detail


Clostridium difficile toxin A-induced apoptosis is p53-independent but depends on glucosylation of Rho GTPases.
MedLine Citation:
PMID:  17437185     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Clostridium difficile toxin A (TcdA) is one of two homologous glucosyltransferases that mono-glucosylate Rho GTPases. HT29 cells were challenged with wild-type and mutant TcdA to investigate the mechanism by which apoptosis is induced. The TcdA-induced re-organization of the actin cytoskeleton led to an increased number of cells within the G2/M phase. Depolymerization of the actin filaments with subsequent G2/M arrest, however, was not causative for apoptosis, as shown in a comparative study using latrunculin B. The activation of caspase-3, -8, and -9 strictly depended on the glucosylation of Rho GTPases. Apoptosis measured by flow cytometry was completely abolished by a pan-caspase inhibitor (z-VAD-fmk). Interestingly, cleavage of procaspase-3 and Bid was not inhibited by z-VAD-fmk, but was inhibited by the calpain/cathepsin inhibitor ALLM. Cleavage of procaspase-8 was susceptible to inhibition by z-VAD-fmk and to the caspase-3 inhibitor Ac-DMQD-CHO, indicating a contribution to the activation of caspase-3 in an amplifying manner. Although TcdA induced mitochondrial damage and cytochrome c release, p53 was not activated or up-regulated. A p53-independent apoptotic effect was also checked by treatment of HCT 116 p53(-/-) cells. In summary, TcdA-induced apoptosis in HT29 cells depends on glucosylation of Rho GTPases leading to activation of cathepsins and caspase-3.
Authors:
Stefanie Nottrott; Janett Schoentaube; Harald Genth; Ingo Just; Ralf Gerhard
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Apoptosis : an international journal on programmed cell death     Volume:  12     ISSN:  1360-8185     ISO Abbreviation:  Apoptosis     Publication Date:  2007 Aug 
Date Detail:
Created Date:  2007-06-25     Completed Date:  2007-11-08     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9712129     Medline TA:  Apoptosis     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1443-53     Citation Subset:  IM    
Affiliation:
Institute of Toxicology, Hannover Medical School, 30625 Hannover, Germany.
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MeSH Terms
Descriptor/Qualifier:
Actins / metabolism
Apoptosis / drug effects*
Bacterial Toxins / metabolism*,  pharmacology*
Caspases / metabolism
Cathepsins / antagonists & inhibitors
Cell Cycle / drug effects
Enterotoxins / metabolism*,  pharmacology*
Enzyme Activation / drug effects
Glucosides / metabolism*
Glycosyltransferases / metabolism,  physiology
HT29 Cells
Humans
Leupeptins / pharmacology
Models, Biological
Protein Processing, Post-Translational / drug effects
Tumor Suppressor Protein p53 / physiology*
rho GTP-Binding Proteins / metabolism*
Chemical
Reg. No./Substance:
0/Actins; 0/Bacterial Toxins; 0/Enterotoxins; 0/Glucosides; 0/Leupeptins; 0/Tumor Suppressor Protein p53; 0/tcdA protein, Clostridium difficile; 110044-82-1/acetylleucyl-leucyl-norleucinal; EC 2.4.-/Glycosyltransferases; EC 3.4.-/Cathepsins; EC 3.4.22.-/Caspases; EC 3.6.5.2/rho GTP-Binding Proteins

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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