Two distinct conformational states define the interaction of human RAD51-ATP with single-stranded DNA.
Garcin, Edwige B
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Brouwer, I., Moschetti, T., Candelli, A., Garcin, E. B., Modesti, M., Pellegrini, L., Wuite, G. J., & et al. (2018). Two distinct conformational states define the interaction of human RAD51-ATP with single-stranded DNA.. EMBO J, 37 (7) https://doi.org/10.15252/embj.201798162
An essential mechanism for repairing DNA double-strand breaks is homologous recombination (HR). One of its core catalysts is human RAD51 (hRAD51), which assembles as a helical nucleoprotein filament on single-stranded DNA, promoting DNA-strand exchange. Here, we study the interaction of hRAD51 with single-stranded DNA using a single-molecule approach. We show that ATP-bound hRAD51 filaments can exist in two different states with different contour lengths and with a free-energy difference of ~4 kBT per hRAD51 monomer. Upon ATP hydrolysis, the filaments convert into a disassembly-competent ADP-bound configuration. In agreement with the single-molecule analysis, we demonstrate the presence of two distinct protomer interfaces in the crystal structure of a hRAD51-ATP filament, providing a structural basis for the two conformational states of the filament. Together, our findings provide evidence that hRAD51-ATP filaments can exist in two interconvertible conformational states, which might be functionally relevant for DNA homology recognition and strand exchange.
DNA-Binding Proteins, Nucleoproteins, DNA, DNA, Single-Stranded, Adenosine Triphosphate, Crystallography, X-Ray, DNA Repair, DNA Replication, Molecular Conformation, Models, Molecular, Rad51 Recombinase, DNA Breaks, Double-Stranded, Homologous Recombination
Wellcome Trust (104641/Z/14/Z)
External DOI: https://doi.org/10.15252/embj.201798162
This record's URL: https://www.repository.cam.ac.uk/handle/1810/280455
Attribution 4.0 International
Licence URL: https://creativecommons.org/licenses/by/4.0/