Abstract
As shown here, mitochondria purified from different organs (liver, brain, kidney, spleen and heart) contain both pro-caspase-9 and the processed, mature form of caspase-9. Purified liver mitochondria release mature caspase-9 upon induction of permeability transition in vitro. This is accompanied by a discrete increase in the enzymatic cleavage of pro-caspase-9 substrates. We found that SHEP neuroblastoma cells constitutively contain pre-processed caspase-9 in their mitochondria, using a combination of subcellular fractionation and immunofluorescence with an antibody specific for the processed caspase. This is a cell type-specific phenomenon since HeLa cells mitochondria mainly contain pro-caspase-9 and comparatively little processed caspase-9. Upon introduction of apoptosis, mitochondrial pro-caspase-9 translocates to the cytosol and to the nucleus. This phenomenon is inhibited by transfection with Bcl-2. In synthesis, we report the unexpected finding that mitochondria can contain a pre-processed caspase isoform in non-apoptotic cells. Bcl-2-mediated regulation of mitochondrial membrane permeabilization may contribute to apoptosis control by preventing mitochondrial, pre-processed caspase-9 from interacting with its cytosolic activators.
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Abbreviations
- AIF:
-
apoptosis-inducing factor
- Apaf-1:
-
apoptosis protease activating factor
- CsA:
-
cyclosporin A
- Cyt c:
-
cytochrome c
- Hsp60:
-
heat shock protein 60
- PT:
-
permeability transition
- t-BHP:
-
tert-butyhlhydroperoxide
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Acknowledgements
We thank Dr. Klaus Michael Debatin (University of Ulm, Germany) for Bcl-2-transfected SHEP cells. This work has been supported by a special grant from the Ligue Nationale contre le Cancer, Comité Val de Marne de la Ligue contre le Cancer, as well as grants from ANRS, FRM, and the European Commission QL61-1999-00739 (to G Kroemer). P Costantini received a fellowship from FRM, J-M Bruey from Ligue Nationale Contre la Cancer (comité de Côle d'Or).
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Costantini, P., Bruey, JM., Castedo, M. et al. Pre-processed caspase-9 contained in mitochondria participates in apoptosis. Cell Death Differ 9, 82–88 (2002). https://doi.org/10.1038/sj.cdd.4400932
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DOI: https://doi.org/10.1038/sj.cdd.4400932
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