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Data from: Topological DNA-binding of SMC-like RecN promotes RecA-mediated DNA double-strand break repair

Citation

Keyamura, Kenji; Hishida, Takashi (2021), Data from: Topological DNA-binding of SMC-like RecN promotes RecA-mediated DNA double-strand break repair, Dryad, Dataset, https://doi.org/10.5061/dryad.7p3p8j3

Abstract

Bacterial RecN, closely related to the structural maintenance of chromosomes (SMC) family of proteins, functions in the repair of DNA double-strand breaks (DSBs) by homologous recombination. However, the understanding of how RecN acts in concert with the RecA recombinase to promote DSB repair remains limited. Here, we demonstrated that the purified Escherichia coli RecN protein topologically loads onto both single-stranded DNA (ssDNA) and double-stranded DNA (dsDNA) that has a preference for ssDNA. RecN topologically bound to dsDNA slides off the end of linear dsDNA, but this is prevented by RecA nucleoprotein filaments on ssDNA, thereby allowing RecN to translocate to DSBs. Furthermore, we found that, once RecN is recruited onto ssDNA, it can topologically capture a second dsDNA substrate in an ATP-dependent manner, suggesting a role in synapsis. Indeed, RecN stimulates RecA-mediated D-loop formation and subsequent strand exchange activities. Our findings provide mechanistic insights into the recruitment of RecN to DSBs and sister chromatid interactions by RecN, both of which function in RecA-mediated DSB repair.

Usage Notes

Source data file contains numerical values that were used to plots graphs from Figures.