The Mre11 complex (Mre11–Rad50–Nbs1) is central to chromosomal maintenance and functions in homologous recombination, telomere maintenance and sister chromatid association1,2,3,4,5,6,7. These functions all imply that the linked binding of two DNA substrates occurs, although the molecular basis for this process remains unknown. Here we present a 2.2 Å crystal structure of the Rad50 coiled-coil region that reveals an unexpected dimer interface at the apex of the coiled coils in which pairs of conserved Cys-X-X-Cys motifs form interlocking hooks that bind one Zn2+ ion. Biochemical, X-ray and electron microscopy data indicate that these hooks can join oppositely protruding Rad50 coiled-coil domains to form a flexible bridge of up to 1,200 Å. This suggests a function for the long insertion in the Rad50 ABC-ATPase domain8. The Rad50 hook is functional, because mutations in this motif confer radiation sensitivity in yeast and disrupt binding at the distant Mre11 nuclease interface. These data support an architectural role for the Rad50 coiled coils in forming metal-mediated bridging complexes between two DNA-binding heads. The resulting assemblies have appropriate lengths and conformational properties to link sister chromatids in homologous recombination and DNA ends in non-homologous end-joining.
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We thank D. Turk for MAIN; M. Gehl for atomic absorption spectroscopy analysis and A.-M. Hayes for technical advice. We acknowledge the key facilities and staff of synchrotron beamlines BL9-2 (SSRL) and 5.0.2 (ALS). Key funding was provided by the US Department of Energy (DOE) and NCI (J.A.T. and J.P.C), the American Cancer Society (J.P.C.), the Canadian Institutes of Health Research (L.C.), National Institutes of Health, Human Frontiers Science Program and DOE (J.H.J.P), Skaggs Institute for Chemical Biology (K.P.H.), the National Institutes of Mental Health (C.T.M.) and The Swiss National Science Foundation (J.L.B).
The authors declare that they have no competing financial interests.
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Hopfner, KP., Craig, L., Moncalian, G. et al. The Rad50 zinc-hook is a structure joining Mre11 complexes in DNA recombination and repair. Nature 418, 562–566 (2002). https://doi.org/10.1038/nature00922
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