Abstract
The resolution of joint molecules that link recombining sister chromatids is essential for chromosome segregation. Here, we determine the fate of unresolved recombination intermediates arising in cells lacking two nucleases required for resolution (GEN1 –/– knockout cells depleted of MUS81). We find that intermediates persist until mitosis and form a distinct class of anaphase bridges, which we term homologous recombination ultra-fine bridges (HR-UFBs). HR-UFBs are distinct from replication stress-associated UFBs, which arise at common fragile sites, and from centromeric UFBs. HR-UFBs are processed by BLM helicase to generate single-stranded RPA-coated bridges that are broken during mitosis. In the next cell cycle, DNA breaks activate the DNA damage checkpoint response, and chromosome fusions arise by non-homologous end joining. Consequently, the cells undergo cell cycle delay and massive cell death. These results lead us to present a model detailing how unresolved recombination intermediates can promote DNA damage and chromosomal instability.
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Acknowledgements
We thank members of the West Lab for their help and encouragement, and K. L. Chan (University of Sussex, UK) for sharing unpublished data. This work was supported by the Francis Crick Institute (FC10212), the European Research Council (ERC-ADG-249145 and ERC-ADG-666400) and the Louis-Jeantet Foundation. The Francis Crick Institute receives core funding from Cancer Research UK, the Medical Research Council and the Wellcome Trust. K.F. is the recipient of a fellowship from the Lundbeck Foundation.
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Y.W.C. carried out the experiments and was assisted by K.F with the fibre assays and some of the immunofluorescence experiments. Y.W.C. and S.C.W. designed the project and wrote the manuscript.
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Chan, Y.W., Fugger, K. & West, S.C. Unresolved recombination intermediates lead to ultra-fine anaphase bridges, chromosome breaks and aberrations. Nat Cell Biol 20, 92–103 (2018). https://doi.org/10.1038/s41556-017-0011-1
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DOI: https://doi.org/10.1038/s41556-017-0011-1
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