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dc.contributor.authorShort, Judith Men
dc.contributor.authorLiu, Yangen
dc.contributor.authorChen, Shaoxiaen
dc.contributor.authorSoni, Neeleshen
dc.contributor.authorMadhusudhan, Mallur Sen
dc.contributor.authorShivji, Mahmud KKen
dc.contributor.authorVenkitaraman, Ashoken
dc.date.accessioned2016-10-24T12:29:08Z
dc.date.available2016-10-24T12:29:08Z
dc.date.issued2016-11-02en
dc.identifier.issn0305-1048
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/260880
dc.description.abstractHomologous DNA recombination (HR) by the RAD51 recombinase enables error-free DNA break repair. To execute HR, RAD51 first forms a presynaptic filament on single-stranded (ss) DNA, which catalyses pairing with homologous double-stranded (ds) DNA. Here, we report a structure for the presynaptic human RAD51 filament at 3.5-5.0Å resolution using electron cryo-microscopy. RAD51 encases ssDNA in a helical filament of 103Å pitch, comprising 6.4 protomers per turn, with a rise of 16.1Å and a twist of 56.2°. Inter-protomer distance correlates with rotation of an α-helical region in the core catalytic domain that is juxtaposed to ssDNA, suggesting how the RAD51-DNA interaction modulates protomer spacing and filament pitch. We map Fanconi anaemia-like disease-associated RAD51 mutations, clarifying potential phenotypes. We predict binding sites on the presynaptic filament for two modules present in each BRC repeat of the BRCA2 tumour suppressor, a critical HR mediator. Structural modelling suggests that changes in filament pitch mask or expose one binding site with filament-inhibitory potential, rationalizing the paradoxical ability of the BRC repeats to either stabilize or inhibit filament formation at different steps during HR. Collectively, our findings provide fresh insight into the structural mechanism of HR and its dysregulation in human disease.
dc.description.sponsorshipMedical Research Council [‘MC_UU_12022/1’, ‘MC_UU_12022/8’ to A.R.V.]; Wellcome Trust-DBT India Alliance Senior Fellowship (to M.S.M.). Funding for open access charge: Medical Research Council.
dc.languageengen
dc.language.isoenen
dc.publisherOxford University Press
dc.rightsAttribution 4.0 International*
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en
dc.subjectAdenosine Diphosphateen
dc.subjectBRCA2 Proteinen
dc.subjectBinding Sitesen
dc.subjectCryoelectron Microscopyen
dc.subjectDNA, Single-Strandeden
dc.subjectGenetic Predisposition to Diseaseen
dc.subjectHomologous Recombinationen
dc.subjectHumansen
dc.subjectModels, Molecularen
dc.subjectMolecular Conformationen
dc.subjectMutationen
dc.subjectProtein Bindingen
dc.subjectProtein Interaction Domains and Motifsen
dc.subjectProtein Subunitsen
dc.subjectRad51 Recombinaseen
dc.subjectRec A Recombinasesen
dc.subjectRepetitive Sequences, Amino Aciden
dc.titleHigh-resolution structure of the presynaptic RAD51 filament on single-stranded DNA by electron cryo-microscopy.en
dc.typeArticle
prism.endingPage9030
prism.issueIdentifier19en
prism.publicationDate2016en
prism.publicationNameNucleic Acids Researchen
prism.startingPage9017
prism.volume44en
dc.identifier.doi10.17863/CAM.6046
dcterms.dateAccepted2016-08-24en
rioxxterms.versionofrecord10.1093/nar/gkw783en
rioxxterms.versionVoRen
rioxxterms.licenseref.urihttp://creativecommons.org/licenses/by/4.0/en
rioxxterms.licenseref.startdate2016-11-02en
dc.identifier.eissn1362-4962
rioxxterms.typeJournal Article/Reviewen
pubs.funder-project-idMedical Research Council (MC_UU_12022/1)
pubs.funder-project-idMRC (MC_UU_12022/8)
pubs.funder-project-idMRC (MC_UU_12022/8)
cam.issuedOnline2016-09-05en
dc.identifier.urlhttps://academic.oup.com/nar/article/44/19/9017/2468442en


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Attribution 4.0 International
Except where otherwise noted, this item's licence is described as Attribution 4.0 International