Abstract
QM31 represents a new class of cytoprotective agents that inhibit the formation of the apoptosome, the caspase activation complex composed by Apaf-1, cytochrome c, dATP and caspase-9. Here, we analyzed the cellular effects of QM31, as compared to the prototypic caspase inhibitor Z-VAD-fmk. QM31 was as efficient as Z-VAD-fmk in suppressing caspase-3 activation, and conferred a similar cytoprotective effect. In contrast to Z-VAD-fmk, QM31 inhibited the release of cytochrome c from mitochondria, an unforeseen property that may contribute to its pronounced cytoprotective activity. Moreover, QM31 suppressed the Apaf-1-dependent intra-S-phase DNA damage checkpoint. These results suggest that QM31 can interfere with the two known functions of Apaf-1, namely apoptosome assembly/activation and intra-S-phase cell cycle arrest. Moreover, QM31 can inhibit mitochondrial outer membrane permeabilization, an effect that is independent from its action on Apaf-1.
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Abbreviations
- Apaf-1:
-
Apoptotic protease activating factor 1
- AnnV:
-
Annexin V
- ATM:
-
Ataxia telangiectasia mutated kinase
- ATR:
-
Ataxia telangiectasia and Rad3 related kinase
- CARD:
-
Caspase activation recruitment domain
- CDDP:
-
Cisplatin
- Chk1:
-
Checkpoint kinase 1
- Cyt c :
-
Cytochrome c
- ΔΨm :
-
Mitochondrial transmembrane potential
- DiOC6(3):
-
3,3′ dihexiloxalocarbocyanine iodide
- FBS:
-
Fetal bovine serum
- FITC:
-
Fluorescein isothiocyanate
- IMS:
-
Mitochondrial intermembrane space
- MOMP:
-
Mitochondrial outer membrane permeabilization
- NSCLC:
-
Non-small cell lung cancer
- OM:
-
Mitochondrial outer membrane
- PARCS:
-
Pro-apoptotic protein required for cell survival
- PGA:
-
Poly-l-glutamic acid
- PI:
-
Propidium iodide
- siRNA:
-
Small interfering RNA
- Z-VAD-fmk:
-
Z-Val-Ala-Asp(OMe)-fluoromethylketone
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Acknowledgments
LM and AM are recipients of predoctoral grants from FPI—MICINN and CSIC, respectively. GK is supported by Cancéropôle Ile-de-France, Institut National du Cancer, Fondation de France, Association Laurette Fugain, Cent pour Sang la Vie, Agence National de la Recherche, and the European Commission (Apo-Sys, ChemoRes. Death-Train, RIGHT). MO acknowledges support from GVPRE/2008/275. AM and EPP are supported by grants BIO2007-60066 and CTQ 2005-00995, respectively, from the Spanish Ministry of Science and Innovation (MICINN).
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Mondragón, L., Galluzzi, L., Mouhamad, S. et al. A chemical inhibitor of Apaf-1 exerts mitochondrioprotective functions and interferes with the intra-S-phase DNA damage checkpoint. Apoptosis 14, 182–190 (2009). https://doi.org/10.1007/s10495-008-0310-x
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DOI: https://doi.org/10.1007/s10495-008-0310-x