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Procaspase-2S inhibits procaspase-3 processing and activation, preventing ROCK-1-mediated apoptotic blebbing and body formation in human B lymphoma Namalwa cells

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Abstract

Procaspase-2S has been reported to selectively prevent membrane blebbing and apoptotic body formation in human monocytic-like leukemic U937 cells after etoposide (VP-16) treatment (Droin et al., Oncogene 20. 260–269, 2001). Here, we show that procaspase-2S overexpressed in human B lymphoma Namalwa cells inhibits procaspase-3 processing and activation, preventing cleavage and activation of Rho GTPase-associated ROCK-1 kinase. Failure of ROCK-1 activation in Namalwa cells correlates with a sustained delay in the appearance of membrane blebbing and apoptotic body formation after VP-16 treatment. Reciprocal coimmunoprecipitation experiments indicate that procaspase-2S binds to procaspase-3, but not procaspase-2L and -9 in untreated and VP-16-treated Namalwa cells. These data suggest that procaspase-2S-mediated anti-apoptotic effects are associated with inhibition of the processing and activation of procaspase-3 in VP-16-treated cells.

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Abbreviations

Ac-DEVD-AMC:

acetyl-Asp-Glu-Val-Asp-7-amino-4-methylcoumarin

Ha-tag:

hemagglutinin epitope Tag sequences

His-tag:

histidine Tag sequences

PAK2:

p21-activated kinase

ROCK-1:

Rho-associated kinase 1

SDS-PAGE:

sodium dodecyl sulfate-polyacrylamide gel electrophoresis

VP-16:

etoposide

z-VDVAD-AFC:

benzyloxycarbonyl-Val-Asp-Val-Ala-Asp-7-amino-4-trifluoromethylcoumarin.

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Author notes

  1. N. Droin

    Present address: LaJolla Institute for Allergy and immunology, San Diego, CA

Authors and Affiliations

  1. Hôpital Notre-Dame and Institut du Cancer de Montréal, Centre de Recherche Centre Hospitalier de l’Université of Montréal (CHUM), Canada

    N. Parent, A.-T. Sané, N. Droin & R. Bertrand

  2. Département de Médecine, Université de Montréal, Montréal (Qué.), Canada

    R. Bertrand

  3. CR-CHUM, Hôpital Notre-Dame and Institut du Cancer de Montréal, 1560 Sherbrooke St. East (Room Y-5634), Montreal (Que.), Canada, H2L 4M1

    R. Bertrand

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  1. N. Parent
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Correspondence to R. Bertrand.

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Parent, N., Sané, AT., Droin, N. et al. Procaspase-2S inhibits procaspase-3 processing and activation, preventing ROCK-1-mediated apoptotic blebbing and body formation in human B lymphoma Namalwa cells. Apoptosis 10, 313–322 (2005). https://doi.org/10.1007/s10495-005-0805-7

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