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PMID: 9922454 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Ordering the cytochrome c-initiated caspase cascade: hierarchical activation of caspases-2, -3, -6, -7, -8, and -10 in a caspase-9-dependent manner.

The Journal of cell biology ·Vol. 144 ·No. 2 ·1999-01-25 ·Pages 281-92

Slee EA, Harte MT, Kluck RM, Wolf BB, Casiano CA, Newmeyer DD, Wang HG, Reed JC, Nicholson DW, Alnemri ES, Green DR, Martin SJ

Abstract

Exit of cytochrome c from mitochondria into the cytosol has been implicated as an important step in apoptosis. In the cytosol, cytochrome c binds to the CED-4 homologue, Apaf-1, thereby triggering Apaf-1-mediated activation of caspase-9. Caspase-9 is thought to propagate the death signal by triggering other caspase activation events, the details of which remain obscure. Here, we report that six additional caspases (caspases-2, -3, -6, -7, -8, and -10) are processed in cell-free extracts in response to cytochrome c, and that three others (caspases-1, -4, and -5) failed to be activated under the same conditions. In vitro association assays confirmed that caspase-9 selectively bound to Apaf-1, whereas caspases-1, -2, -3, -6, -7, -8, and -10 did not. Depletion of caspase-9 from cell extracts abrogated cytochrome c-inducible activation of caspases-2, -3, -6, -7, -8, and -10, suggesting that caspase-9 is required for all of these downstream caspase activation events. Immunodepletion of caspases-3, -6, and -7 from cell extracts enabled us to order the sequence of caspase activation events downstream of caspase-9 and reveal the presence of a branched caspase cascade. Caspase-3 is required for the activation of four other caspases (-2, -6, -8, and -10) in this pathway and also participates in a feedback amplification loop involving caspase-9.

MeSH Terms
Animals Apoptosis Apoptotic Protease-Activating Factor 1 Caspase 10 Caspase 2 Caspase 3 Caspase 6 Caspase 7 Caspase 8 Caspase 9 Caspases/metabolism Cell Extracts Cytochrome c Group/metabolism Enzyme Activation Humans Jurkat Cells Protein Processing, Post-Translational Proteins/metabolism Rabbits
Chemicals
APAF1 protein, human Apoptotic Protease-Activating Factor 1 Cell Extracts Cytochrome c Group Proteins CASP3 protein, human CASP6 protein, human CASP7 protein, human CASP8 protein, human CASP9 protein, human Caspase 10 Caspase 2 Caspase 3 Caspase 6 Caspase 7 Caspase 8 Caspase 9 Caspases CASP10 protein, human
Authors & Affiliations
12 authors, click to expand affiliations / ORCID
Slee E A
Molecular Cell Biology Laboratory, Department of Biology, National University of Ireland, Maynooth, Co. Kildare, Ireland.
Harte M T
Kluck R M
Wolf B B
Casiano C A
Newmeyer D D
Wang H G
Reed J C
Nicholson D W
Alnemri E S
Green D R
Martin S J
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.144.2.281
Published
1999-01-25
Pages
281-92
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2132895
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.144.2.281 PMC Full Text View journal details
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