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Nbs1 is essential for DNA repair by homologous recombination in higher vertebrate cells

Nature volume 420, pages 93–98 (2002)Cite this article

Abstract

Double-strand breaks occur during DNA replication and are also induced by ionizing radiation. There are at least two pathways which can repair such breaks: non-homologous end joining and homologous recombination (HR). Although these pathways are essentially independent of one another, it is possible that the proteins Mre11, Rad50 and Xrs2 are involved in both pathways in Saccharomyces cerevisiae1. In vertebrate cells, little is known about the exact function of the Mre11–Rad50–Nbs1 complex in the repair of double-strand breaks because Mre11- and Rad50-null mutations are lethal2. Here we show that Nbs1 is essential for HR-mediated repair in higher vertebrate cells. The disruption of Nbs1 reduces gene conversion and sister chromatid exchanges, similar to other HR-deficient mutants3. In fact, a site-specific double-strand break repair assay showed a notable reduction of HR events following generation of such breaks in Nbs1-disrupted cells. The rare recombinants observed in the Nbs1-disrupted cells were frequently found to have aberrant structures, which possibly arise from unusual crossover events, suggesting that the Nbs1 complex might be required to process recombination intermediates.

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Figure 1: Generation of Nbs1-/-/- clones.
Figure 2: Cellular phenotypes of Nbs1-/-/- cells.
Figure 3: Recombination of the SCneo reporter construct in Nbs1-/-/- cells.

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Acknowledgements

We thank L. N. Kapp for helping to prepare the manuscript; M. Jasin for providing the SCneo construct; A. Shinohara for anti-Rad51 antibody. We also thank H. Komata, C. Muranaka, T. Jo, M. Ueda, A. Kodama-Kamesako and A. Okamoto for laboratory assistance. This work was supported in part by the Ministry of Education, Science, Sports and Culture of Japan (H.T. and K.K.), and by the Nuclear Safety Research Association of Japan (H.T.).

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Authors and Affiliations

  1. Department of Environmental Sciences, Faculty of Science, Ibaraki University, Bunkyo 2-1-1, Mito, Ibaraki, 310-8512, Japan

    Hiroshi Tauchi

  2. Department of Radiation Biology, Research Institute for Radiation Biology and Medicine, Hiroshima University, Kasumi 1-2-3, 734-8553, Minami-ku, Hiroshima, Japan

    Junya Kobayashi, Ken-ichi Morishima, Takahiro Shiraishi, Emi Ito, Asako Nakamura, Shinya Matsuura & Kenshi Komatsu

  3. Department of Cell Biology and Genetics, Erasmus University Rotterdam, PO Box 1738, 3000 DR, Rotterdam, The Netherlands

    Dik C. van Gent, Nicole S. Verkaik & Diana vanHeems

  4. Department of Radiation Genetics, Faculty of Medicine, Radiation Biology Research Center, Kyoto University, 606-8501, Yoshida Konoe, Sakyo-ku, Kyoto, Japan

    Kenshi Komatsu

  5. Immunology and Molecular Genetics, Kawasaki Medical School, 577 Matsushima, 701-0192, Kurashiki, Okayama, Japan

    Minoru Takata

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Tauchi, H., Kobayashi, J., Morishima, Ki. et al. Nbs1 is essential for DNA repair by homologous recombination in higher vertebrate cells. Nature 420, 93–98 (2002). https://doi.org/10.1038/nature01125

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