Post-replicative repair involves separase-dependent removal of the kleisin subunit of cohesin

Abstract

DNA double-strand break repair is critical for cell viability and involves highly coordinated pathways to restore DNA integrity at the lesion. An early event during homology-dependent repair is resection of the break to generate progressively longer 3′ single-strand tails that are used to identify suitable templates for repair1. Sister chromatids provide near-perfect sequence homology and are therefore the preferred templates during homologous recombination2,3. To provide a bias for the use of sisters as donors, cohesin—the complex that tethers sister chromatids together4—is recruited to the break5,6 to enforce physical proximity. Here we show that DNA breaks promote dissociation of cohesin loaded during the previous S phase in budding yeast, and that damage-induced dissociation of cohesin requires separase, the protease that dissolves cohesion in anaphase7. Moreover, a separase-resistant allele of the gene coding for the α-kleisin subunit of cohesin, Mcd1 (also known as Scc1), reduces double-strand break resection and compromises the efficiency of repair even when loaded during DNA damage. We conclude that post-replicative DNA repair involves cohesin dissociation by separase to promote accessibility to repair factors during the coordinated cellular response to restore DNA integrity.

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Figure 1: S-phase-loaded cohesin is not sufficient to provide cohesion of a chromatin fragment in the presence of DNA damage.
Figure 2: Mcd1 dissociation in the presence of DNA damage requires separase.
Figure 3: Mcd1 dissociation facilitates 5′–3′ end resection.
Figure 4: Mcd1 dissociation facilitates repair of DNA DSB.

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Acknowledgements

We would like to thank D. Koshland, E. Unal, J. Haber, G. Ira and K. Nasmyth for providing valuable strains. We thank members of the Aragon laboratory, P. Ullal, S. Farmer, K. Pegram, J. Torres-Rosell and M. Merkenschlager for discussions and reading of the manuscript. This work was funded by the Intramural Research Program of the Medical Research Council UK.

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Contributions

A.M. made the original observation that transcriptional repression of MCD1 in the presence of DNA damage causes defects in sister chromatid cohesion. A.M. and A.C.-B. performed all ChIP analysis. A.M., A.C.-B., N.S. and V.C.-P. performed microscopic analysis of cohesion. A.C.-B. and M.E. performed physical assays to measure resection and repair. A.M. and A.C.-B. performed biochemical assays to measure Mcd1 stability. A.J. provided technical support. L.A. wrote the paper, and all authors analysed the data, discussed the results and commented on the manuscript.

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Correspondence to Luis Aragón.

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The authors declare no competing financial interests.

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McAleenan, A., Clemente-Blanco, A., Cordon-Preciado, V. et al. Post-replicative repair involves separase-dependent removal of the kleisin subunit of cohesin. Nature 493, 250–254 (2013). https://doi.org/10.1038/nature11630

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