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  • Original Paper
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Ku antigen is required to relieve G2 arrest caused by inhibition of DNA topoisomerase II activity by the bisdioxopiperazine ICRF-193

Oncogene volume 20, pages 1990–1999 (2001)Cite this article

Abstract

Ku antigen is necessary for DNA double-strand break (DSB) repair through its ability to bind DNA ends with high affinity and to recruit the catalytic subunit of DNA-PK to the DSBs. Ku-deficient cells are hypersensitive to agents causing DSBs in DNA but also to the DNA topoisomerase II (topo II) inhibitor ICRF-193, which does not induce DSBs. This suggests a new role of Ku antigen, that is independent of DSB repair by DNA-PK. Here we characterize the basis for the hypersensitivity of Ku-deficient cells to ICRF-193. Chromosome condensation and segregation, which are dependent on topo II, but also the catalytic activity of topo II in late S-G2 were inhibited to a comparable extent when ICRF-193 was applied to Ku-deficient cells or wild-type cells. However, mutant cells arrested in G2 by ICRF-193 treatment were unable to progress into M phase upon drug removal, although drug-trapped topo II complexes were removed from DNA and the two isoforms of topo II recovered their catalytic activity as in wild-type cells. The reversibility of G2 arrest was recovered by complementation of mutant cells with a human Ku86 cDNA. Notably, chromosome condensation was abnormal in Ku-deficient cells after suppression of the G2 arrest by caffeine, even in the absence of ICRF-193. These results reflect the involvement of Ku-antigen in the cellular response to topo II inhibition, more particularly in relieving G2 arrest caused by topo II inhibition in late S/G2 and the subsequent recovery of chromosome condensation.

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Acknowledgements

We would like to thank Dr Creighton for the generous gift of ICRF-193, Dr Kikuchi for monoclonal antibodies against DNA topoisomerase IIβ and Dr Blanchard for antibodies against GAPDH. Particular thanks to Drs R Hipskind, E Schwob, J Tazi and I Robbins for their comments on the manuscript and also to Dr JM Blanchard for his continuous interest. This work was supported by the Ministère de la Recherche et de l'Enseignement Supérieur and the Association pour la Recherche contre le Cancer. P Muñoz was supported by a post-doctoral fellowship from the European Community and the Fondation pour la Recherche Médicale.

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  1. Purificación Muñoz

    Present address: Unitat de Bioquimica, Facultat D'Odontologia, Universitat de Barcelona, C/Feixa Llarga s/n, 08907 L'Hospitalet de Llobregat, Barcelona, Spain

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  1. Institut de Génétique Moléculaire de Montpellier, UMR 5535, IFR 24, CNRS, 1919 route de Mende, 34293 Montpellier Cedex 5, France

    Purificación Muñoz, Fabienne Baus & Jacques Piette

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Muñoz, P., Baus, F. & Piette, J. Ku antigen is required to relieve G2 arrest caused by inhibition of DNA topoisomerase II activity by the bisdioxopiperazine ICRF-193. Oncogene 20, 1990–1999 (2001). https://doi.org/10.1038/sj.onc.1204262

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