Data for manuscript entitled "The dynamic conformational ensemble of dsDNA-bound Mre11-Rad50 supports a structure-function model of its catalytic cycle"

The MRE11-RAD50-NBS1/Xrs2 (MRN/X) protein complex has essential roles in the repair of damaged DNA. The current understanding of the conformational landscape of the core MR complex comes from a multitude of structural studies. However, given the heterogeneous nature of these structures, we suspected that a number of conformational states may still be unaccounted for. Using methyl-based NMR experiments on P. furiosus MR, we determined an ensemble of distinct conformations of MR bound to DNA, consistent with the highly dynamic nature of the MR-DNA complex. Some structural models represented previously solved structures, but others have not been observed before. Interrogation of these structures via in vitro activity assays on MR mutants revealed an unexpected, striking correlation between the nuclease activity of Mre11 and the magnitude of DNA-stimulated ATP hydrolysis by Rad50. The data support a model for MR activity where DNA-stimulated ATP hydrolysis unlocks Rad50 to provide access to the Mre11 active sites and further demonstrate how a heterogeneous ensemble of conformations can be used to coordinate various functions to direct biological outcomes.