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PMID: 11406587 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Direct cleavage of the human DNA fragmentation factor-45 by granzyme B induces caspase-activated DNase release and DNA fragmentation.

The EMBO journal ·Vol. 20 ·No. 12 ·2001-06-15 ·Pages 3101-13

Sharif-Askari E, Alam A, Rhéaume E, Beresford PJ, Scotto C, Sharma K, Lee D, DeWolf WE, Nuttall ME, Lieberman J, Sékaly RP

Abstract

The protease granzyme B (GrB) plays a key role in the cytocidal activity during cytotoxic T lymphocyte (CTL)-mediated programmed cell death. Multiple caspases have been identified as direct substrates for GrB, suggesting that the activation of caspases constitutes an important event during CTL-induced cell death. However, recent studies have provided evidence for caspase-independent pathway(s) during CTL-mediated apoptosis. In this study, we demonstrate caspase-independent and direct cleavage of the 45 kDa unit of DNA fragmentation factor (DFF45) by GrB both in vitro and in vivo. Using a novel and selective caspase-3 inhibitor, we show the ability of GrB to process DFF45 directly and mediate DNA fragmentation in the absence of caspase-3 activity. Furthermore, studies with DFF45 mutants reveal that both caspase-3 and GrB share a common cleavage site, which is necessary and sufficient to induce DNA fragmentation in target cells during apoptosis. Together, our data suggest that CTLs possess alternative mechanism(s) for inducing DNA fragmentation without the requirement for caspases.

MeSH Terms
Apoptosis Apoptosis Regulatory Proteins Binding Sites Caspase 3 Caspase Inhibitors Caspases/metabolism DNA Fragmentation Deoxyribonucleases/metabolism Enzyme Activation Granzymes Humans Jurkat Cells Molecular Structure Nuclear Proteins/chemistry,metabolism Proteins/chemistry,metabolism Serine Endopeptidases/metabolism Substrate Specificity
Chemicals
Apoptosis Regulatory Proteins Caspase Inhibitors Nuclear Proteins Proteins caspase-activated DNase inhibitor Deoxyribonucleases GZMB protein, human Granzymes Serine Endopeptidases CASP3 protein, human Caspase 3 Caspases
Authors & Affiliations
11 authors, click to expand affiliations / ORCID
Sharif-Askari E
Laboratoire d'Immunologie, Département de Microbiologie et d'Immunologie, Université de Montréal, Montréal, H3C 3J7.
Alam A
Rhéaume E
Beresford P J
Scotto C
Sharma K
Lee D
DeWolf W E
Nuttall M E
Lieberman J
Sékaly R P
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/20.12.3101
Published
2001-06-15
Pages
3101-13
Language
English
Region
England
NLM ID
8208664
PMCID
PMC150191
Subset
IM
Grants
NIAID NIH HHS · R01 AI045587 · United States
NIAID NIH HHS · R56 AI045587 · United States
NIAID NIH HHS · AI 45587 · United States
External Links
PubMed source DOI: 10.1093/emboj/20.12.3101 PMC Full Text View journal details
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