Architectural plasticity of human BRCA2-RAD51 complexes in DNA break repair.
The tumor suppressor BRCA2 is a large multifunctional protein mutated in 50-60% of familial breast cancers. BRCA2 interacts with many partners and includes multiple regions with potentially disordered structure. In homology directed DNA repair BRCA2 delivers RAD51 to DNA resulting in removal of RPA and assembly of a RAD51 nucleoprotein filament. Dynamic rearrangements of BRCA2 likely drive this molecular hand-off initiating DNA strand exchange. We show human BRCA2 forms oligomers which can have an extended shape. Scanning force microscopy and quantitative single molecule fluorescence define the variety of BRCA2 complexes, reveal dramatic rearrangements upon RAD51 binding and the loading of RAD51 patches on single strand DNA. At sites of repair in cell nuclei, super-resolution microscopy shows BRCA2 and RAD51 arranged in largely separate locations. We identified dynamic structural transitions in BRCA2 complexes suggested to facilitate loading of RAD51 onto RPA coated single strand DNA and subsequent release of BRCA2.
Department of Molecular Genetics, Cancer Genomics Center Netherlands, Erasmus University Medical Center, 3000 CA Rotterdam, The Netherlands.
Humberto
H
Sánchez
Erasmus Optical Imaging Centre, Erasmus University Medical Center, 3000 CA Rotterdam, The Netherlands.|Department of Pathology, Erasmus University Medical Center, 3000 CA Rotterdam, The Netherlands.
Maarten W
MW
Paul
Department of Molecular Genetics, Cancer Genomics Center Netherlands, Erasmus University Medical Center, 3000 CA Rotterdam, The Netherlands.
Malgorzata
M
Grosbart
Department of Molecular Genetics, Cancer Genomics Center Netherlands, Erasmus University Medical Center, 3000 CA Rotterdam, The Netherlands.|Department of Radiation Oncology, Erasmus University Medical Center, 3000 CA Rotterdam, The Netherlands.
Sarah E
SE
van Rossum-Fikkert
Department of Molecular Genetics, Cancer Genomics Center Netherlands, Erasmus University Medical Center, 3000 CA Rotterdam, The Netherlands.|Department of Radiation Oncology, Erasmus University Medical Center, 3000 CA Rotterdam, The Netherlands.
Joyce H G
JHG
Lebbink
Department of Molecular Genetics, Cancer Genomics Center Netherlands, Erasmus University Medical Center, 3000 CA Rotterdam, The Netherlands.|Department of Radiation Oncology, Erasmus University Medical Center, 3000 CA Rotterdam, The Netherlands.
Roland
R
Kanaar
Erasmus Optical Imaging Centre, Erasmus University Medical Center, 3000 CA Rotterdam, The Netherlands.|Department of Pathology, Erasmus University Medical Center, 3000 CA Rotterdam, The Netherlands.
Adriaan B
AB
Houtsmuller
Department of Molecular Genetics, Cancer Genomics Center Netherlands, Erasmus University Medical Center, 3000 CA Rotterdam, The Netherlands.|Department of Radiation Oncology, Erasmus University Medical Center, 3000 CA Rotterdam, The Netherlands.
Claire
C
Wyman
D024682:BRCA2 Protein / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D001665:Binding Sites; D002467:Cell Nucleus / Q000235:genetics* / Q000378:metabolism / Q000648:ultrastructure; D053904:DNA Breaks, Single-Stranded; D004277:DNA, Single-Stranded / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D064267:Intrinsically Disordered Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D018625:Microscopy, Atomic Force; D011485:Protein Binding; D055503:Protein Multimerization; D059767:Recombinational DNA Repair; D051716:Replication Protein A / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D000072760:Single Molecule Imaging
D024682:BRCA2 Protein; C551750:BRCA2 protein, human; D004277:DNA, Single-Stranded; D004268:DNA-Binding Proteins; D064267:Intrinsically Disordered Proteins; C448305:RAD51C protein, human; D051716:Replication Protein A