Document Detail


A serine protease is involved in the initiation of DNA damage-induced apoptosis.
MedLine Citation:
PMID:  14502243     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Caspases are considered to be the key effector proteases of apoptosis. Initiator caspases cleave and activate downstream executioner caspases, which are responsible for the degradation of numerous cellular substrates. We studied the role of caspases in apoptotic cell death of a human melanoma cell line. Surprisingly, the pancaspase inhibitor zVAD-fmk was unable to block cleavage of poly(ADP-ribose) polymerase (PARP) after treatment with etoposide, while it did prevent DEVDase activity. It is highly unlikely that caspase-2, which is a relatively zVAD-fmk-resistant caspase, is mediating etoposide-induced PARP cleavage, as a preferred inhibitor of this caspase could not prevent cleavage. In contrast, caspase activation and PARP degradation were blocked by pretreatment of the cells with the serine protease inhibitor 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF). We therefore conclude that a serine protease regulates an alternative initiation mechanism that leads to caspase activation and PARP cleavage. More importantly, while zVAD-fmk could not rescue melanoma cells from etoposide-induced death, the combination with AEBSF resulted in substantial protection. This indicates that this novel pathway fulfills a critical role in the execution of etoposide-induced programmed cell death.
Authors:
E C de Bruin; D Meersma; J de Wilde; I den Otter; E M Schipper; J P Medema; L T C Peltenburg
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Cell death and differentiation     Volume:  10     ISSN:  1350-9047     ISO Abbreviation:  Cell Death Differ.     Publication Date:  2003 Oct 
Date Detail:
Created Date:  2003-09-22     Completed Date:  2004-06-07     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  9437445     Medline TA:  Cell Death Differ     Country:  England    
Other Details:
Languages:  eng     Pagination:  1204-12     Citation Subset:  IM    
Affiliation:
Department of Clinical Oncology, Leiden University Medical Center, Leiden, The Netherlands.
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MeSH Terms
Descriptor/Qualifier:
Amino Acid Chloromethyl Ketones / pharmacology
Animals
Apoptosis / drug effects,  genetics*
Blotting, Western
Caspase 2
Caspase 3
Caspases / antagonists & inhibitors,  metabolism
Cell Line
Cell Line, Tumor / drug effects,  radiation effects
Coumarins / metabolism
Cysteine Proteinase Inhibitors / pharmacology
DNA Damage*
Etoposide / pharmacology
Fibroblasts / cytology,  drug effects,  metabolism
Flow Cytometry
Humans
Microscopy, Phase-Contrast
Oligopeptides / metabolism,  pharmacology
Peptide Hydrolases / metabolism
Poly(ADP-ribose) Polymerases / metabolism
Rats
Serine Endopeptidases / metabolism*
Serine Proteinase Inhibitors / pharmacology
Sulfones / pharmacology
Tumor Necrosis Factor-alpha / pharmacology
Chemical
Reg. No./Substance:
0/Amino Acid Chloromethyl Ketones; 0/Coumarins; 0/Cysteine Proteinase Inhibitors; 0/Oligopeptides; 0/Serine Proteinase Inhibitors; 0/Sulfones; 0/Tumor Necrosis Factor-alpha; 0/aspartyl-glutamyl-valyl-aspartyl-7-amino-4-trifluoromethylcoumarin; 0/benzoylcarbonyl-valyl-aspartyl-valyl-alanyl-aspartyl-fluoromethyl ketone; 0/benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone; 33419-42-0/Etoposide; 34284-75-8/4-(2-aminoethyl)benzenesulfonylfluoride; EC 2.4.2.30/Poly(ADP-ribose) Polymerases; EC 3.4.-/Peptide Hydrolases; EC 3.4.21.-/Serine Endopeptidases; EC 3.4.22.-/CASP3 protein, human; EC 3.4.22.-/Casp3 protein, rat; EC 3.4.22.-/Caspase 2; EC 3.4.22.-/Caspase 3; EC 3.4.22.-/Caspases; EC 3.4.99.-/DEVDase

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