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dc.contributor.authorHollfelder, Florianen
dc.contributor.authorLindenburg, Laurensen
dc.contributor.authorHyvonen, Markoen
dc.contributor.authorKaminski, Clemensen
dc.contributor.authorRees, Ericen
dc.contributor.authorGielen, Fabriceen
dc.contributor.authorButz, Marenen
dc.contributor.authorDowns, Jessicaen
dc.contributor.authorPantelejevs, Teodorsen
dc.contributor.authorZuazua-Villar, Pedroen
dc.date.accessioned2021-08-18T15:00:03Z
dc.date.available2021-08-18T15:00:03Z
dc.identifier.issn0027-8424
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/326909
dc.description.abstractExchanges of protein sequence modules support leaps in function unavailable through point mutations during evolution. Here we study the role of the two RAD51-interacting modules within the eight binding BRC repeats of BRCA2. We created 64 chimeric repeats by shuffling these modules and measured their binding to RAD51. We found that certain shuffled module combinations were stronger binders than any of the module combinations in the natural repeats. Surprisingly, the contribution from the two modules was poorly correlated with affinities of natural repeats, with a weak BRC8 repeat containing the most effective N-terminal module. The binding of the strongest chimera, BRC8-2, to RAD51 was improved by -2.4 kCal/mol compared to the strongest natural repeat, BRC4. A crystal structure of RAD51:BRC8-2 complex shows an improved interface fit and an extended b-hairpin in this repeat. BRC8-2 was shown to function in human cells, preventing the formation of nuclear RAD51 foci after ionizing radiation.
dc.description.sponsorshipBBSRC (BB/K013629/1), Cancer Research UK (C7905/A25715), EPSRC (grants EP/L015889/1 and EP/H018301/1), the Wellcome Trust (grants 3-3249/Z/16/Z and 089703/Z/09/Z), MRC (grants MR/K015850/1 and MR/K02292X/1), ERC, H2020 Marie-Curie
dc.publisherNational Academy of Sciences
dc.rightsAll rights reserved
dc.titleImproved RAD51 binders through motif shuffling based on the modularity of BRC repeatsen
dc.typeArticle
prism.publicationNameProceedings of the National Academy of Sciences of USAen
dc.identifier.doi10.17863/CAM.74358
dcterms.dateAccepted2021-08-11en
rioxxterms.versionAM
rioxxterms.licenseref.urihttp://www.rioxx.net/licenses/all-rights-reserveden
rioxxterms.licenseref.startdate2021-08-11en
dc.contributor.orcidHollfelder, Florian [0000-0002-1367-6312]
dc.contributor.orcidHyvonen, Marko [0000-0001-8683-4070]
rioxxterms.typeJournal Article/Reviewen
pubs.funder-project-idBBSRC (BB/K013629/1)
cam.orpheus.counter10*
rioxxterms.freetoread.startdate2024-08-18


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