Imperial College London
Portrait Picture

Professor David S. Rueda

Faculty of Medicine, Department of Infectious Disease

Chair in Molecular and Cellular Biophysics
 
 
 
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Contact

 

david.rueda Website

 
 
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Location

 

6.12DLMS BuildingHammersmith Campus

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Summary

 

Publications

Citation

BibTex format

@article{Belan:2023:10.1016/j.molcel.2023.06.031,
author = {Belan, O and Greenhough, L and Kuhlen, L and Anand, R and Kaczmarczyk, A and Gruszka, DT and Yardimci, H and Zhang, X and Rueda, DS and West, SC and Boulton, SJ},
doi = {10.1016/j.molcel.2023.06.031},
journal = {Molecular Cell},
pages = {2925--2940.e8},
title = {Visualization of direct and diffusion-assisted RAD51 nucleation by full-length human BRCA2 protein},
url = {http://dx.doi.org/10.1016/j.molcel.2023.06.031},
volume = {83},
year = {2023}
}
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RIS format (EndNote, RefMan)

TY  - JOUR
AB  - 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.
AU  - Belan,O
AU  - Greenhough,L
AU  - Kuhlen,L
AU  - Anand,R
AU  - Kaczmarczyk,A
AU  - Gruszka,DT
AU  - Yardimci,H
AU  - Zhang,X
AU  - Rueda,DS
AU  - West,SC
AU  - Boulton,SJ
DO  - 10.1016/j.molcel.2023.06.031
EP  - 2940
PY  - 2023///
SN  - 1097-2765
SP  - 2925
TI  - Visualization of direct and diffusion-assisted RAD51 nucleation by full-length human BRCA2 protein
T2  - Molecular Cell
UR  - http://dx.doi.org/10.1016/j.molcel.2023.06.031
UR  - http://hdl.handle.net/10044/1/105797
VL  - 83
ER  -
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