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Nuclear colocalization and interaction between bcl-xL and cdk1(cdc2) during G2/M cell-cycle checkpoint

Oncogene volume 26, pages 5851–5865 (2007)Cite this article

Abstract

In response to cancer chemotherapeutic drugs, cells rapidly trigger the apoptotic program or undergo growth arrest and senescence at specific phases of the cell cycle. Mitochondrial bcl-xL plays a central role in preventing alteration of mitochondrial dysfunction, cytochrome c release, caspase activation, DNA fragmentation and apoptosis. However, its pleitropic function depends on its subcellular localization. Here, we show that in addition to its mitochondrial effect that delays apoptosis, bcl-xL colocalizes and binds to cdk1(cdc2) during G2/M cell-cycle checkpoint and its overexpression stabilizes a G2/M-arrest senescence program in surviving cells after DNA damage. Bcl-xL potently inhibits cdk1(cdc2) kinase activity, which is reversible by a synthetic peptide between the 41st amino acid and 60th amino acid surrounding of the Thr47 and Ser62 phosphorylation sites, and Asn52 deamidation site, within the flexible loop domain of bcl-xL. A mutant deleted of this region does not alter the antiapoptotic function of bcl-xL, but impedes its effect on cdk1(cdc2) activity and on the G2/M-arrest senescence program after DNA damage. The nuclear interaction of bcl-xL and cdk1(cdc2) suggests that bcl-xL is coupled to the stabilization of a cell-cycle checkpoint induced by DNA damage, and this effect is genetically distinct from its function on apoptosis.

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Abbreviations

Ac-DEVD-AMC:

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

cdk(cdc):

cyclin-dependent kinase

cisPT:

cis-platinum (II) diamine dichloride

CPT:

20-S-camptothecin lactone

DEVDase:

Asp-Glu-Val-Asp protease

HA-tag:

hemagglutinin epitope tag sequences

PCR:

polymerase chain reaction

SDS–PAGE:

sodium dodecyl sulfate–polyacrylamide gel electrophoresis

TNF-α:

tumor necrosis factor alpha

VP-16:

etoposide

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Acknowledgements

This work was initiated by a grant from the National Cancer Institute of Canada to RB, and completed under a grant from the Cancer Research Society Inc. to RB. ES obtained a fellowship from the Fonds de la recherche en santé du Québec. The editorial work of Mr Ovid Da Silva is much appreciated.

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  1. Centre de recherche, Centre hospitalier de l'Université of Montréal – Hôpital Notre-Dame and Institut du Cancer de Montréal, Montréal, Quebec, Canada

    E Schmitt, M Beauchemin & R Bertrand

  2. Departement de Médecine, Université de Montréal, Montréal, Quebec, Canada

    R Bertrand

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Correspondence to R Bertrand.

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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc).

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Schmitt, E., Beauchemin, M. & Bertrand, R. Nuclear colocalization and interaction between bcl-xL and cdk1(cdc2) during G2/M cell-cycle checkpoint. Oncogene 26, 5851–5865 (2007). https://doi.org/10.1038/sj.onc.1210396

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