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

Published version
Peer-reviewed

Type

Article

Change log

Authors

Brouwer, Ineke 
Moschetti, Tommaso 
Candelli, Andrea 
Garcin, Edwige B 
Modesti, Mauro 

Abstract

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.

Description

Keywords

DNA repair, RAD51, homologous recombination, single‐stranded DNA, Adenosine Triphosphate, Crystallography, X-Ray, DNA, DNA Breaks, Double-Stranded, DNA Repair, DNA Replication, DNA, Single-Stranded, DNA-Binding Proteins, Homologous Recombination, Models, Molecular, Molecular Conformation, Nucleoproteins, Rad51 Recombinase

Journal Title

EMBO J

Conference Name

Journal ISSN

0261-4189
1460-2075

Volume Title

37

Publisher

Springer Science and Business Media LLC
Sponsorship
Wellcome Trust (104641/Z/14/Z)