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Meiotic instability of human minisatellite CEB1 in yeast requires DNA double-strand breaks

Nature Genetics volume 23, pages 367–371 (1999)Cite this article

Abstract

Minisatellites are tandemly repeated DNA sequences of 10–100-bp units1. Some minisatellite loci are highly unstable in the human germ line, and structural analysis of mutant alleles has suggested that repeat instability results from a recombination-based process. To provide insights into the molecular mechanism of human minisatellite instability, we developed Saccharomyces cerevisiae strains carrying alleles of the most unstable human minisatellite locus, CEB1 (ref. 2). We observed that CEB1 is destabilized in meiosis, resulting in a variety of intra- and inter-allelic gains or losses of repeat units, similar to rearrangements described in humans3. Using mutations affecting the initiation of recombination4,5 (spo11) or mismatch repair6 (msh2 pms1 ), we demonstrate that meiotic destabilization depends on the initiation of homologous recombination at nearby DNA double-strand break (DSBs) sites and involves a 'rearranged heteroduplex' intermediate. Most of the human and yeast data can be explained and unified in the context of DSB repair models7.

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Figure 1: Experimental system.
Figure 2: Detection of meiotic double-strand breaks near the CEB1 insert on chromosome VIII.
Figure 3: Examples of intra- and inter-allelic CEB1 rearrangements obtained in the msh2 pms1 background.
Figure 4: A DSB-repair model to explain meiotic CEB1 rearrangements.

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Acknowledgements

We thank members of our laboratories for technical advice, materials and helpful discussions; K. Smith for English correction; and R. Rothstein for helpful discussion. This study was supported by an initial grant from the Groupement de Recherches et d'Etudes sur les Génomes and subsequently by the Association de Recherche contre le Cancer (ARC), the Ligue Nationale contre le Cancer, the D.G.A/D.S.P/S.T.T.C, the Human Frontier Scientific Program and the Radiobiology program of the Institut Curie. H.D. was supported by graduate student fellowships from the MNERT and the ARC.

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Authors and Affiliations

  1. Institut Curie, Section de Recherche, CNRS UMR144, Paris, France

    Hélène Debrauwère & Alain Nicolas

  2. Institut de Biologie, Nantes, France

    Jérôme Buard, Jacques Tessier, Dominique Aubert & Gilles Vergnaud

  3. Institut de Génétique et de Microbiologie, Université Paris-Sud, Orsay, France

    Jacques Tessier & Gilles Vergnaud

  4. Centre d'Etudes du Bouchet, Vert le Petit , France

    Gilles Vergnaud

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  1. Hélène Debrauwère
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  2. Jérôme Buard
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  4. Dominique Aubert
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  5. Gilles Vergnaud
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  6. Alain Nicolas
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Correspondence to Alain Nicolas.

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Debrauwère, H., Buard, J., Tessier, J. et al. Meiotic instability of human minisatellite CEB1 in yeast requires DNA double-strand breaks. Nat Genet 23, 367–371 (1999). https://doi.org/10.1038/15557

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