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A chemical inhibitor of Apaf-1 exerts mitochondrioprotective functions and interferes with the intra-S-phase DNA damage checkpoint

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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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Authors and Affiliations

  1. U848, INSERM, 94805, Villejuif, France

    Laura Mondragón, Lorenzo Galluzzi, Shahul Mouhamad, José-Miguel Vicencio, Ilio Vitale & Guido Kroemer

  2. Institut Gustave Roussy, PR1, 39 rue Camille Desmoulins, 94805, Villejuif, France

    Laura Mondragón, Lorenzo Galluzzi, Shahul Mouhamad, José-Miguel Vicencio, Ilio Vitale & Guido Kroemer

  3. Université Paris-Sud/Paris XI, 94270, Le Kremlin Bicêtre, France

    Laura Mondragón, Lorenzo Galluzzi, Shahul Mouhamad, José-Miguel Vicencio, Ilio Vitale & Guido Kroemer

  4. Department of Medicinal Chemistry, Centro de Investigacion Principe Felipe, 46013, Valencia, Spain

    Laura Mondragón, Mar Orzáez & Enrique Perez-Paya

  5. U841, INSERM, 94010, Créteil, France

    Shahul Mouhamad

  6. Hôpital Henri Mondor, 94010, Créteil, France

    Shahul Mouhamad

  7. Department of Chemical and Molecular Nanotechnology, IQAC-CSIC, 08034, Barcelona, Spain

    Alejandra Moure & Angel Messeguer

  8. Instituto de Biomedicina de Valencia, CSIC, 46010, Valencia, Spain

    Enrique Perez-Paya

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  1. Laura Mondragón
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  2. Lorenzo Galluzzi
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  9. Enrique Perez-Paya
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  10. Guido Kroemer
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Correspondence to Guido Kroemer.

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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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