Document Detail


Internucleosomal DNA cleavage in apoptotic WEHI 231 cells is mediated by a chymotrypsin-like protease.
MedLine Citation:
PMID:  15507121     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Although several lines of evidence support a role for serine proteases in apoptosis, little is known about the mechanisms involved. In the present study, we have examined the apoptosis-inducing potential and dissected the death-signalling pathways of N-tosyl-L-phenylalanine chloromethyl ketone (TPCK) and N-tosyl-L-lysine chloromethyl ketone (TLCK), inhibitors of chymotrypsin- and trypsin-like proteases, respectively. Our results designate two distinct roles for serine proteases. Firstly, we show that both inhibitors induce biochemical and morphological characteristics of apoptosis, including proteolysis of poly(ADP-ribose) polymerase 1 (PARP-1) and inhibitor of caspase-activated DNase (ICAD), as well as mitochondrial dysfunction, and that their action is abrogated by the caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp.fluoromethylketone (z-VAD.fmk). These results suggest that inhibition of anti-apoptotic serine proteases governs the onset of the caspase-dependant apoptotic cascade. Secondly, we also demonstrate the involvement of a serine protease in the terminal stage of apoptosis. We showed that chymotrypsin-like protease activity is required for internucleosomal DNA fragmentation in apoptotic cells. Hence, DNA fragmentation is abrogated in TPCK-pre-treated WEHI 231 cells undergoing apoptosis triggered either by anti-IgM or TLCK. These results indicate that internucleosomal DNA cleavage in apoptotic cells is mediated by a chymotrypsin-like protease.
Authors:
Jernej Murn; Uros Urleb; Irena Mlinaric-Rascan
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Genes to cells : devoted to molecular & cellular mechanisms     Volume:  9     ISSN:  1356-9597     ISO Abbreviation:  Genes Cells     Publication Date:  2004 Nov 
Date Detail:
Created Date:  2004-10-27     Completed Date:  2005-04-11     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  9607379     Medline TA:  Genes Cells     Country:  England    
Other Details:
Languages:  eng     Pagination:  1103-11     Citation Subset:  IM    
Affiliation:
Faculty of Pharmacy, University of Ljubljana, Askerceva 7, 1000 Ljubljana, Slovenia.
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MeSH Terms
Descriptor/Qualifier:
Apoptosis*
Blotting, Western
Caspases / metabolism
Cell Line
DNA / metabolism*
Enzyme Activation
Flow Cytometry
Hydrolysis
Membrane Potentials
Permeability
Serine Endopeptidases / metabolism*
Chemical
Reg. No./Substance:
9007-49-2/DNA; EC 3.4.21.-/Serine Endopeptidases; EC 3.4.22.-/Caspases

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


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