A RAD51–ADP double filament structure unveils the mechanism of filament dynamics in homologous recombination

ATP-dependent RAD51 recombinases play an essential role in eukaryotic homologous recombination by catalyzing a four-step process: 1) formation of a RAD51 single-filament assembly on ssDNA in the presence of ATP, 2) complementary DNA strand-exchange, 3) ATP hydrolysis transforming the RAD51 filament...

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Published in:Nature communications 2023-08, Vol.14 (1), p.4993-4993, Article 4993
Main Authors: Luo, Shih-Chi, Yeh, Min-Chi, Lien, Yu-Hsiang, Yeh, Hsin-Yi, Siao, Huei-Lun, Tu, I-Ping, Chi, Peter, Ho, Meng-Chiao
Format: Article
Language:English
Subjects:
101/28
631/337/1427/2190
631/535/1258/1259
Adenosine diphosphate
ATP
Deoxyribonucleic acid
Dismantling
DNA
DNA structure
Filaments
Homologous recombination
Homology
Humanities and Social Sciences
Intermediates
multidisciplinary
Packaging
Proteins
Science
Science (multidisciplinary)
Structural analysis
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Summary:ATP-dependent RAD51 recombinases play an essential role in eukaryotic homologous recombination by catalyzing a four-step process: 1) formation of a RAD51 single-filament assembly on ssDNA in the presence of ATP, 2) complementary DNA strand-exchange, 3) ATP hydrolysis transforming the RAD51 filament into an ADP-bound disassembly-competent state, and 4) RAD51 disassembly to provide access for DNA repairing enzymes. Of these steps, filament dynamics between the ATP- and ADP-bound states, and the RAD51 disassembly mechanism, are poorly understood due to the lack of near-atomic-resolution information of the ADP-bound RAD51–DNA filament structure. We report the cryo-EM structure of ADP-bound RAD51–DNA filaments at 3.1 Å resolution, revealing a unique RAD51 double-filament that wraps around ssDNA. Structural analysis, supported by ATP-chase and time-resolved cryo-EM experiments, reveals a collapsing mechanism involving two four-protomer movements along ssDNA for mechanical transition between RAD51 single- and double-filament without RAD51 dissociation. This mechanism enables elastic change of RAD51 filament length during structural transitions between ATP- and ADP-states. RAD51 filaments are essential for eukaryotic homologous recombination. This study shows the cryo-EM structure of an ADP-bound RAD51 double-filament that wraps around ssDNA revealing a stepwise collapsing mechanism for the transition between the ATP/ADP-bound filament intermediates.
ISSN:2041-1723
2041-1723
DOI:10.1038/s41467-023-40672-5