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Modulation of apoptosis by procaspase-2 short isoform: selective inhibition of chromatin condensation, apoptotic body formation and phosphatidylserine externalization

Oncogene volume 20, pages 260–269 (2001)Cite this article

Abstract

Procaspase-2 is one of the cysteine aspartate proteases involved in apoptotic cell death. Alternative splicing of CASP-2 messenger RNA generates a long isoform, procaspase-2L, whose overexpression induces cell death and a truncated isoform, procaspase-2S, whose function remains poorly defined. The present study explored the consequences of procaspase-2S overexpression in U937 human leukemic cells exposed to the topoisomerase II inhibitor etoposide as an apoptotic stimulus. Overexpression of procaspase-2S in U937 cells partially prevented nuclear changes associated with etoposide-induced cell death, as determined by Hoechst 33342 staining of nuclear chromatin and electron microscopy studies. Procaspase-2S also prevented the maturation of apoptotic bodies, delayed phosphatidylserine externalization on the plasma membrane and prevented the cleavage and activation of procaspase-2L. These effects were not observed when the cysteine 289 in the consensus QACRG motif was mutated into a serine. Wild-type procaspase-2S overexpression did not influence the cleavage of procaspase-3, procaspase-7 and poly(ADP-ribose)polymerase nor the fragmentation of nuclear DNA into nucleosome-sized fragments. Altogether, these results indicate that the short isoform of procaspase-2 negatively interferes with selective features of apoptosis, an activity that is suppressed by mutation of the cysteine 289.

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Abbreviations

AIF:

apoptosis-inducing factor

CAD:

caspase-activated DNAse

CARD:

caspase recrutment domain

CRADD:

caspase and RIP adaptator with death domain

ICAD:

inhibitor of CAD

PCR:

polymerase chain reaction

RAIDD:

RIP-associated ICH-1/CED-3-homologous protein with a death domain

z-VDVAD-AFC:

benzyloxycarbonyl-Valine-Aspartate-Valine-Alanine-Aspartate-7-amino-4-trifluoromethyl coumarin)

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Acknowledgements

The authors thank Dr Nicholson (Merck, Toronto, Canada) for kindly providing us an anti-caspase-3 Ab. This work was supported in part by grants from the Medical Research Council of Canada (MT-15019) to R Bertrand while E Solary's research group is labeled and granted by the Ligue Nationale Contre le Cancer. R Bertrand is a scholar from the Medical Research Council of Canada and the Cancer Research Society Inc (Canada). N Droin received supports from the Ministère de l'Education Nationale, de l'Enseignement et de la Recherche, the Société Française d'Hématologie and the French Ministère des Affaires Etrangères.

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Author notes

  1. Richard Bertrand and Eric Solary: R Bertrand and E Solary are the senior authors and contributed equally to this study

Authors and Affiliations

  1. INSERM U517, Faculties of Medicine & Pharmacy, 7 boulevard Jeanne d'Arc, Dijon, 21033, France

    Nathalie Droin, Cedric Rébé, Florence Bichat, Arlette Hammann & Eric Solary

  2. Research Centre of University of Montreal Hospital Centre, Notre Dame Hospital, 1560 Sherbrooke Street East, Montreal, H2L 4M1, Quebec, Canada

    Nathalie Droin & Richard Bertrand

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Droin, N., Rébé, C., Bichat, F. et al. Modulation of apoptosis by procaspase-2 short isoform: selective inhibition of chromatin condensation, apoptotic body formation and phosphatidylserine externalization. Oncogene 20, 260–269 (2001). https://doi.org/10.1038/sj.onc.1204066

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