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Single-molecule analysis reveals cooperative stimulation of Rad51 filament nucleation and growth by mediator proteins.
Belan, Ondrej; Barroso, Consuelo; Kaczmarczyk, Artur; Anand, Roopesh; Federico, Stefania; O'Reilly, Nicola; Newton, Matthew D; Maeots, Erik; Enchev, Radoslav I; Martinez-Perez, Enrique; Rueda, David S; Boulton, Simon J.
Afiliação
  • Belan O; DSB Repair Metabolism Laboratory, The Francis Crick Institute, London NW1 1AT, UK.
  • Barroso C; Meiosis group, MRC-London Institute of Medical Sciences, London W12 0NN, UK.
  • 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.
  • Anand R; DSB Repair Metabolism Laboratory, The Francis Crick Institute, London NW1 1AT, UK.
  • Federico S; Peptide Chemistry STP, The Francis Crick Institute, London NW1 1AT, UK.
  • O'Reilly N; Peptide Chemistry STP, The Francis Crick Institute, London NW1 1AT, UK.
  • 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.
  • Maeots E; Visual Biochemistry Laboratory, The Francis Crick Institute, London NW1 1AT, UK.
  • Enchev RI; Visual Biochemistry Laboratory, The Francis Crick Institute, London NW1 1AT, UK.
  • Martinez-Perez E; Meiosis group, MRC-London Institute of Medical Sciences, London W12 0NN, UK.
  • 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.
  • Boulton SJ; DSB Repair Metabolism Laboratory, The Francis Crick Institute, London NW1 1AT, UK. Electronic address: simon.boulton@crick.ac.uk.
Mol Cell ; 81(5): 1058-1073.e7, 2021 03 04.
Article em En | MEDLINE | ID: mdl-33421363
ABSTRACT
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.
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Texto completo: 1 Base de dados: MEDLINE Assunto principal: Proteínas de Transporte / Caenorhabditis elegans / DNA de Helmintos / Proteínas de Caenorhabditis elegans / Proteínas de Ligação a DNA / Rad51 Recombinase / Reparo de DNA por Recombinação Limite: Animals Idioma: En Ano de publicação: 2021 Tipo de documento: Article
Texto completo
Imprimir
XML
PubMed Links
Buscar no Google

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Proteínas de Transporte / Caenorhabditis elegans / DNA de Helmintos / Proteínas de Caenorhabditis elegans / Proteínas de Ligação a DNA / Rad51 Recombinase / Reparo de DNA por Recombinação Limite: Animals Idioma: En Ano de publicação: 2021 Tipo de documento: Article