Two distinct conformational states define the interaction of human RAD51‐ATP with single‐stranded DNA

• Ineke Brouwer ^[1] ; Tommaso Moschetti ^[2] ; Andrea Candelli ^[1] ; Edwige B Garcin ^[3] ; Mauro Modesti ^[3] ; Luca Pellegrini ^[2] ; Gijs JL Wuite ^[1] ; Erwin JG Peterman ^[1]
  1. [1] VU University Amsterdam

     VU University Amsterdam

     Países Bajos

     
  2. [2] University of Cambridge

     University of Cambridge

     Cambridge District, Reino Unido

     
  3. [3] 3 Cancer Research Center of Marseille CNRS UMR7258 Inserm U1068 Institut Paoli‐Calmettes Aix‐Marseille Université Marseille France

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• Localización: EMBO journal: European Molecular Biology Organization, ISSN 0261-4189, Vol. 37, Nº. 7, 2018
• Idioma: inglés
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  • 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.