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Two distinct conformational states define the interaction of human RAD51-ATP with single-stranded DNA.

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cam.issuedOnline2018-03-05
dc.contributor.authorBrouwer, Ineke
dc.contributor.authorMoschetti, Tommaso
dc.contributor.authorCandelli, Andrea
dc.contributor.authorGarcin, Edwige B
dc.contributor.authorModesti, Mauro
dc.contributor.authorPellegrini, Luca
dc.contributor.authorWuite, Gijs Jl
dc.contributor.authorPeterman, Erwin Jg
dc.contributor.orcidPellegrini, Luca [0000-0002-9300-497X]
dc.contributor.orcidWuite, Gijs Jl [0000-0002-5706-043X]
dc.contributor.orcidPeterman, Erwin Jg [0000-0003-1058-249X]
dc.date.accessioned2018-09-20T12:03:52Z
dc.date.available2018-09-20T12:03:52Z
dc.date.issued2018-04-03
dc.description.abstractAn essential mechanism for repairing DNA double-strand breaks is homologous recombination (HR). One of its core catalysts is human RAD51 (hRAD51), which assembles as a helical nucleoprotein filament on single-stranded DNA, promoting DNA-strand exchange. Here, we study the interaction of hRAD51 with single-stranded DNA using a single-molecule approach. We show that ATP-bound hRAD51 filaments can exist in two different states with different contour lengths and with a free-energy difference of ~4 kBT per hRAD51 monomer. Upon ATP hydrolysis, the filaments convert into a disassembly-competent ADP-bound configuration. In agreement with the single-molecule analysis, we demonstrate the presence of two distinct protomer interfaces in the crystal structure of a hRAD51-ATP filament, providing a structural basis for the two conformational states of the filament. Together, our findings provide evidence that hRAD51-ATP filaments can exist in two interconvertible conformational states, which might be functionally relevant for DNA homology recognition and strand exchange.
dc.format.mediumPrint-Electronic
dc.identifier.doi10.17863/CAM.27826
dc.identifier.eissn1460-2075
dc.identifier.issn0261-4189
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/280455
dc.languageeng
dc.language.isoeng
dc.publisherSpringer Science and Business Media LLC
dc.publisher.urlhttp://dx.doi.org/10.15252/embj.201798162
dc.rightsAttribution 4.0 International
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/
dc.subjectDNA repair
dc.subjectRAD51
dc.subjecthomologous recombination
dc.subjectsingle‐stranded DNA
dc.subjectAdenosine Triphosphate
dc.subjectCrystallography, X-Ray
dc.subjectDNA
dc.subjectDNA Breaks, Double-Stranded
dc.subjectDNA Repair
dc.subjectDNA Replication
dc.subjectDNA, Single-Stranded
dc.subjectDNA-Binding Proteins
dc.subjectHomologous Recombination
dc.subjectModels, Molecular
dc.subjectMolecular Conformation
dc.subjectNucleoproteins
dc.subjectRad51 Recombinase
dc.titleTwo distinct conformational states define the interaction of human RAD51-ATP with single-stranded DNA.
dc.typeArticle
dcterms.dateAccepted2018-02-08
prism.issueIdentifier7
prism.publicationDate2018
prism.publicationNameEMBO J
prism.volume37
pubs.funder-project-idWellcome Trust (104641/Z/14/Z)
rioxxterms.licenseref.startdate2018-04
rioxxterms.licenseref.urihttp://www.rioxx.net/licenses/all-rights-reserved
rioxxterms.typeJournal Article/Review
rioxxterms.versionVoR
rioxxterms.versionofrecord10.15252/embj.201798162

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