Single-molecule analysis reveals cooperative stimulation of Rad51 filament nucleation and growth by mediator proteins.

Animals Caenorhabditis elegans / genetics Caenorhabditis elegans Proteins / genetics Carrier Proteins / genetics DNA Breaks, Double-Stranded DNA, Helminth / genetics DNA-Binding Proteins / genetics Gene Expression Regulation Molecular Chaperones / genetics Mutation Protein Binding Rad51 Recombinase / genetics Recombinational DNA Repair Replication Protein A / genetics Signal Transduction Single Molecule Imaging
BRCA2 DNA repair Rad51 nucleoprotein filaments Rad51 paralogs homologous recombination single molecule approaches

Journal

Molecular cell
ISSN: 1097-4164
Titre abrégé: Mol Cell
Pays: United States
ID NLM: 9802571

Informations de publication

Date de publication:
04 03 2021
Historique:
received: 06 08 2020
revised: 02 11 2020
accepted: 10 12 2020
pubmed: 10 1 2021
medline: 17 3 2021
entrez: 9 1 2021
Statut: ppublish

Résumé

Homologous recombination (HR) is an essential DNA double-strand break (DSB) repair mechanism, which is frequently inactivated in cancer. During HR, RAD51 forms nucleoprotein filaments on RPA-coated, resected DNA and catalyzes strand invasion into homologous duplex DNA. How RAD51 displaces RPA and assembles into long HR-proficient filaments remains uncertain. Here, we employed single-molecule imaging to investigate the mechanism of nematode RAD-51 filament growth in the presence of BRC-2 (BRCA2) and RAD-51 paralogs, RFS-1/RIP-1. BRC-2 nucleates RAD-51 on RPA-coated DNA, whereas RFS-1/RIP-1 acts as a "chaperone" to promote 3' to 5' filament growth via highly dynamic engagement with 5' filament ends. Inhibiting ATPase or mutation in the RFS-1 Walker box leads to RFS-1/RIP-1 retention on RAD-51 filaments and hinders growth. The rfs-1 Walker box mutants display sensitivity to DNA damage and accumulate RAD-51 complexes non-functional for HR in vivo. Our work reveals the mechanism of RAD-51 nucleation and filament growth in the presence of recombination mediators.

Identifiants

DOI: 10.1016/j.molcel.2020.12.020 PMID: 33421363 PMC: PMC7941204
pubmed: 33421363
pii: S1097-2765(20)30939-4
doi: 10.1016/j.molcel.2020.12.020
pmc: PMC7941204
pii:
doi:

Substances chimiques

BRC-2 protein, C elegans 0
Caenorhabditis elegans Proteins 0
Carrier Proteins 0
DNA, Helminth 0
DNA-Binding Proteins 0
Molecular Chaperones 0
RFS-1 protein, C elegans 0
RIP-1 protein, C elegans 0
Replication Protein A 0
Rad51 Recombinase EC 2.7.7.-
rad-51 protein, C elegans EC 2.7.7.-

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

1058-1073.e7

Subventions

Organisme : Medical Research Council
ID : FC0010048
Pays : United Kingdom
Organisme : Wellcome Trust
ID : 206292/Z/17/Z
Pays : United Kingdom
Organisme : Medical Research Council
ID : M-A652-5PY60
Pays : United Kingdom
Organisme : Medical Research Council
ID : MC_UP_1102/5
Pays : United Kingdom
Organisme : Cancer Research UK
ID : FC0010048
Pays : United Kingdom
Organisme : Medical Research Council
ID : MC_U120097113
Pays : United Kingdom
Organisme : Wellcome Trust
ID : FC0010048
Pays : United Kingdom
Organisme : Wellcome Trust
Pays : United Kingdom
Organisme : Medical Research Council
ID : MC-A658-5TY10
Pays : United Kingdom
Organisme : Wellcome Trust
ID : P67153
Pays : United Kingdom

Commentaires et corrections

Type : CommentIn

Informations de copyright

Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.

Déclaration de conflit d'intérêts

Declaration of interests S.J.B. is also scientific co-founder and VP Science Strategy at Artios Pharma Ltd., Babraham Research Campus, Cambridge, UK. The other authors declare no competing interests.

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Auteurs

Ondrej Belan (O)

DSB Repair Metabolism Laboratory, The Francis Crick Institute, London NW1 1AT, UK.

Consuelo Barroso (C)

Meiosis group, MRC-London Institute of Medical Sciences, London W12 0NN, UK.

Artur Kaczmarczyk (A)

Department of Infectious Disease, Faculty of Medicine, Imperial College London, London W12 0NN, UK; Single Molecule Imaging Group, MRC-London Institute of Medical Sciences, London W12 0NN, UK.

Roopesh Anand (R)

DSB Repair Metabolism Laboratory, The Francis Crick Institute, London NW1 1AT, UK.

Stefania Federico (S)

Peptide Chemistry STP, The Francis Crick Institute, London NW1 1AT, UK.

Nicola O'Reilly (N)

Peptide Chemistry STP, The Francis Crick Institute, London NW1 1AT, UK.

Matthew D Newton (MD)

Department of Infectious Disease, Faculty of Medicine, Imperial College London, London W12 0NN, UK; Single Molecule Imaging Group, MRC-London Institute of Medical Sciences, London W12 0NN, UK.

Erik Maeots (E)

Visual Biochemistry Laboratory, The Francis Crick Institute, London NW1 1AT, UK.

Radoslav I Enchev (RI)

Visual Biochemistry Laboratory, The Francis Crick Institute, London NW1 1AT, UK.

Enrique Martinez-Perez (E)

Meiosis group, MRC-London Institute of Medical Sciences, London W12 0NN, UK.

David S Rueda (DS)

Department of Infectious Disease, Faculty of Medicine, Imperial College London, London W12 0NN, UK; Single Molecule Imaging Group, MRC-London Institute of Medical Sciences, London W12 0NN, UK. Electronic address: david.rueda@imperial.ac.uk.

Simon J Boulton (SJ)

DSB Repair Metabolism Laboratory, The Francis Crick Institute, London NW1 1AT, UK. Electronic address: simon.boulton@crick.ac.uk.

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Classifications MeSH

questionsmedicales.fr › Eucaryotes › Animaux › Single-molecule analysis reveals cooperative...
questionsmedicales.fr › Eucaryotes › Animaux › Invertébrés › Helminthes › Nematoda › Chromadorea › Rhabditida › Rhabditoidea › Caenorhabditis › Caenorhabditis elegans › Single-molecule analysis reveals cooperative...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines d'helminthes › Protéines de Caenorhabditis elegans › Single-molecule analysis reveals cooperative...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines de transport › Single-molecule analysis reveals cooperative...
questionsmedicales.fr › Phénomènes génétiques › Altération de l'ADN › Cassures de l'ADN › Cassures double-brin de l'ADN › Single-molecule analysis reveals cooperative...
questionsmedicales.fr › Acides nucléiques, nucléotides et nucléosides › Acides nucléiques › ADN › ADN des helminthes › Single-molecule analysis reveals cooperative...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines de liaison à l'ADN › Single-molecule analysis reveals cooperative...
questionsmedicales.fr › Phénomènes génétiques › Régulation de l'expression des gènes › Single-molecule analysis reveals cooperative...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Chaperons moléculaires › Single-molecule analysis reveals cooperative...
questionsmedicales.fr › Phénomènes génétiques › Variation génétique › Mutation › Single-molecule analysis reveals cooperative...
questionsmedicales.fr › Métabolisme › Liaison aux protéines › Single-molecule analysis reveals cooperative...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines des groupes de complémentation de l'anémie de Fanconi › Rad51 Recombinase › Single-molecule analysis reveals cooperative...
questionsmedicales.fr › Phénomènes génétiques › Recombinaison génétique › Recombinaison homologue › Réparation de l'ADN par recombinaison › Single-molecule analysis reveals cooperative...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines de liaison à l'ADN › Protéine A de réplication › Single-molecule analysis reveals cooperative...
questionsmedicales.fr › Phénomènes physiologiques cellulaires › Transduction du signal › Single-molecule analysis reveals cooperative...
questionsmedicales.fr › Techniques d'investigation › Techniques de sonde moléculaire › Imagerie moléculaire › Imagerie de molécules uniques › Single-molecule analysis reveals cooperative...