The Mre11–Rad50–Nbs1 (MRN) complex tethers, processes and signals DNA double-strand breaks, promoting genomic stability. To understand the functional architecture of MRN, we determined the crystal structures of the Schizosaccharomyces pombe Mre11 dimeric catalytic domain alone and in complex with a fragment of Nbs1. Two Nbs1 subunits stretch around the outside of the nuclease domains of Mre11, with one subunit additionally bridging and locking the Mre11 dimer via a highly conserved asymmetrical binding motif. Our results show that Mre11 forms a flexible dimer and suggest that Nbs1 not only is a checkpoint adaptor but also functionally influences Mre11-Rad50. Clinical mutations in Mre11 are located along the Nbs1-interaction sites and weaken the Mre11-Nbs1 interaction. However, they differentially affect DNA repair and telomere maintenance in Saccharomyces cerevisiae, potentially providing insight into their different human disease pathologies.
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We are grateful to J. Petrini (Memorial Sloan-Kettering Cancer Center, New York) for his gift of antibodies to Mre11, Rad50 and Xrs2; members of the Hopfner lab for technical support and discussions; and M. Bennett, A. Rojowska, A. Kopetzki and C. Jung for help with experimentation. We thank the Max Planck Institute crystallization facility for crystallization trials, the staffs of the synchrotron beamlines for help with data collection and processing and SLS and ESRF for beamtime allowance. Research in the K.-P.H. lab was funded by grants from the German Research Council (SFBs 684, 646 and TR5), the German Excellence Initiative, European Commission (IP DNA repair), and US National Institutes of Health (U19AI83025). Research in the K.S. lab was funded by grants from the German Research Council (SFB 646) and the European Research Council (ERC; ERC Starting Grant, project 204522). Research in the S.P.J. lab is supported by grants from Cancer Research UK (C6/A11226), the European Research Council, the European Community's Seventh Framework Program (FP7/2007-2013) under grant agreement HEALTH-F2-2010-259893 and by core infrastructure funding from Cancer Research UK and the Wellcome Trust. S.P.J. receives his salary from the University of Cambridge, supplemented by Cancer Research UK.
The authors declare no competing financial interests.
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Schiller, C., Lammens, K., Guerini, I. et al. Structure of Mre11–Nbs1 complex yields insights into ataxia-telangiectasia–like disease mutations and DNA damage signaling. Nat Struct Mol Biol 19, 693–700 (2012). https://doi.org/10.1038/nsmb.2323
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