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DNA damage induce γ-tubulin–RAD51 nuclear complexes in mammalian cells

Oncogene volume 24, pages 5165–5172 (2005)Cite this article

Abstract

Rad51 protein plays an essential role in recombination repair of DNA double-strand breaks and DNA crosslinking adducts. It is part of complexes which can vary with the stage of the cell cycle and the nature of the DNA lesions. During a search for Rad51-associated proteins in CHO nuclear extracts of S-phase cells by mass spectrometry of proteins immunoprecipitated with Rad51 antibodies, we identified a centrosomal protein, γ-tubulin. This association was confirmed by the reverse immunoprecipitation with γ-tubulin antibodies. Both proteins copurified from HeLa cells nuclear extracts following a tandem affinity purification of double-tagged Rad51. Immunofluorescence analysis showed colocalization of both Rad51 and γ-tubulin in discrete foci in mammalian cell nuclei. The number of colocalized foci and their overlapping area increased in the presence of DNA damage produced by genotoxic treatments either during S phase or in exponentially growing cells. These variations did not result from an overall stress because microtubule cytoskeleton poisons devoid of direct interactions with DNA, such as taxol or colcemid, did not lead to an increase of this association. The recruitment of Rad51 and γ-tubulin in the same nuclear complex suggests a link between DNA recombination repair and the centrosome function during the cell cycle.

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Acknowledgements

We gratefully acknowledge Dr NP Johnson for critical reading of the manuscript, A Moisand for EM and F Viala for iconography. This work was supported by a grant from Ligue régionale de recherche sur le cancer Midi-Pyrénées (to MD), a grant from the Association de recherche contre le cancer (MW) and a grant from the ARECA network of Association de recherche contre le cancer to MD, MW and BM and from Région Midi-Pyrénées (BM).

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Authors and Affiliations

  1. IPBS, UMR 5089, CNRS Université Paul Sabatier, 205 Route de Narbonne, 31077, Toulouse Cedex, 4, France

    Claire Lesca, Maryse Germanier, Carole Pichereaux, Stéphane Emond, Odile Burlet-Schiltz, Bernard Monsarrat & Martine Defais

  2. ISTMT, CRPS, UMR2587, CNRS-Pierre Fabre, 3 rue des Satellites, Toulouse, 31400, France

    Brigitte Raynaud-Messina, Chantal Etievant & Michel Wright

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  1. Claire Lesca
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Correspondence to Martine Defais.

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Lesca, C., Germanier, M., Raynaud-Messina, B. et al. DNA damage induce γ-tubulin–RAD51 nuclear complexes in mammalian cells. Oncogene 24, 5165–5172 (2005). https://doi.org/10.1038/sj.onc.1208723

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