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Induction of multiple double-strand breaks within an hsr by meganucleaseI-SceI expression or fragile site activation leads to formation of double minutes and other chromosomal rearrangements

Oncogene volume 21, pages 7671–7679 (2002)Cite this article

Abstract

Gene amplification is frequently associated with tumor progression, hence, understanding the underlying mechanisms is important. The study of in vitro model systems indicated that different initial mechanisms accumulate amplified copies within the chromosomes (hsr) or on extra-chromosomal elements (dmin). It has long been suggested that formation of dmin could also occur following hsr breakdown. In order to check this hypothesis, we developed an approach based on the properties of the I-SceI meganuclease, which induces targeted DNA double-strand breaks. A clone containing an I-SceI site, integrated by chance close to an endogenous dhfr gene locus, was used to select for methotrexate resistant mutants. We recovered clones in which the I-SceI site was passively co-amplified with the dhfr gene within the same hsr. We show that I-SceI-induced hsr breakdown leads to the formation of dmin and creates different types of chromosomal rearrangements, including inversions. This demonstrates, for the first time, a direct relationship between double-strand breaks and inversions. Finally, we show that activation of fragile sites by aphidicolin or hypoxia in hsr-containing cells also generates dmin and a variety of chromosomal rearrangements. This may constitute a valuable model to study the consequences of breaks induced in hsr of cancer cells in vivo.

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Acknowledgements

We thank Drs M Weiss and G Buttin for careful reading of the manuscript and helpful discussion, B Dujon for providing the I-SceI gene and the protein recognition sequence, J Hamlin for the cosmids of the dhfr locus. This work was supported in part by the Association pour la Recherche sur le Cancer, the Fondation de France, the Ligue Nationale Française contre le Cancer (Comité de Paris). A Coquelle is supported by a fellowship of the Ligue Nationale Contre le Cancer and L Rozier is a Fellow of the MENRT.

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  1. Unité de Cytogénétique Moléculaire et Oncologie (UMR 147 CNRS), Institut Curie, 26 rue d'Ulm, Paris, Cédex 05, 75248, France

    Arnaud Coquelle, Lorène Rozier, Bernard Dutrillaux & Michelle Debatisse

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  1. Arnaud Coquelle
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  2. Lorène Rozier
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Correspondence to Michelle Debatisse.

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Coquelle, A., Rozier, L., Dutrillaux, B. et al. Induction of multiple double-strand breaks within an hsr by meganucleaseI-SceI expression or fragile site activation leads to formation of double minutes and other chromosomal rearrangements. Oncogene 21, 7671–7679 (2002). https://doi.org/10.1038/sj.onc.1205880

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