Visualization of direct and diffusion-assisted RAD51 nucleation by full-length human BRCA2 protein.
BRCA2
DNA damage and repair
Rad51 filament formation
homologous recombination
single-molecule analysis
tumor suppressor protein
Journal
Molecular cell
ISSN: 1097-4164
Titre abrégé: Mol Cell
Pays: United States
ID NLM: 9802571
Informations de publication
Date de publication:
17 08 2023
17 08 2023
Historique:
received:
03
02
2023
revised:
09
06
2023
accepted:
26
06
2023
medline:
21
8
2023
pubmed:
28
7
2023
entrez:
27
7
2023
Statut:
ppublish
Résumé
Homologous recombination (HR) is essential for error-free repair of DNA double-strand breaks, perturbed replication forks (RFs), and post-replicative single-stranded DNA (ssDNA) gaps. To initiate HR, the recombination mediator and tumor suppressor protein BRCA2 facilitates nucleation of RAD51 on ssDNA prior to stimulation of RAD51 filament growth by RAD51 paralogs. Although ssDNA binding by BRCA2 has been implicated in RAD51 nucleation, the function of double-stranded DNA (dsDNA) binding by BRCA2 remains unclear. Here, we exploit single-molecule (SM) imaging to visualize BRCA2-mediated RAD51 nucleation in real time using purified proteins. We report that BRCA2 nucleates and stabilizes RAD51 on ssDNA either directly or through an unappreciated diffusion-assisted delivery mechanism involving binding to and sliding along dsDNA, which requires the cooperative action of multiple dsDNA-binding modules in BRCA2. Collectively, our work reveals two distinct mechanisms of BRCA2-dependent RAD51 loading onto ssDNA, which we propose are critical for its diverse functions in maintaining genome stability and cancer suppression.
Identifiants
pubmed: 37499663
pii: S1097-2765(23)00507-5
doi: 10.1016/j.molcel.2023.06.031
pmc: PMC7615647
mid: EMS193943
pii:
doi:
Substances chimiques
BRCA2 Protein
0
Rad51 Recombinase
EC 2.7.7.-
DNA-Binding Proteins
0
DNA, Single-Stranded
0
DNA
9007-49-2
RAD51 protein, human
EC 2.7.7.-
BRCA2 protein, human
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
2925-2940.e8Subventions
Organisme : Wellcome Trust
ID : 206292/Z/17/Z
Pays : United Kingdom
Organisme : Medical Research Council
ID : MC_UP_1102/5
Pays : United Kingdom
Organisme : Wellcome Trust
ID : 210658
Pays : United Kingdom
Organisme : Medical Research Council
ID : CC2057
Pays : United Kingdom
Organisme : Cancer Research UK
ID : CC2057
Pays : United Kingdom
Organisme : Wellcome Trust
Pays : United Kingdom
Organisme : Wellcome Trust
ID : CC2057
Pays : United Kingdom
Informations de copyright
Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.
Déclaration de conflit d'intérêts
Declaration of interests S.J.B. is a co-founder, VP Science Strategy, and a shareholder of Artios Pharma Ltd. S.J.B. is also a member of the advisory board of Molecular Cell.
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