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TitleA RAD51-ADP double filament structure unveils the mechanism of filament dynamics in homologous recombination.
Journal, issue, pagesNat Commun, Vol. 14, Issue 1, Page 4993, Year 2023
Publish dateAug 17, 2023
AuthorsShih-Chi Luo / Min-Chi Yeh / Yu-Hsiang Lien / Hsin-Yi Yeh / Huei-Lun Siao / I-Ping Tu / Peter Chi / Meng-Chiao Ho /
PubMed AbstractATP-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 ...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.
External linksNat Commun / PubMed:37591853 / PubMed Central
MethodsEM (helical sym.)
Resolution3.14 - 3.58 Å
Structure data

34372

8gyk

EMDB-34372, PDB-8gyk:
CryoEM structure of the RAD51_ADP filament
Method: EM (helical sym.) / Resolution: 3.14 Å

EMDB-36040: Composite map of the Rad51-ADP filament
Method: EM (helical sym.) / Resolution: 3.58 Å

Chemicals

ChemComp-ADP:
ADENOSINE-5'-DIPHOSPHATE / ADP, energy-carrying molecule*YM

ChemComp-MG:
Unknown entry

Source
  • homo sapiens (human)
KeywordsRECOMBINATION / Recombinase / ATPase / DNA repair protein / DNA binding

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