Show simple item record

dc.contributor.authorCole, Danielen
dc.contributor.authorRajendra, Eesonen
dc.contributor.authorRoberts-Thomson, Meredithen
dc.contributor.authorHardwick, Brynen
dc.contributor.authorMcKenzie, Grahame Jen
dc.contributor.authorPayne, Michaelen
dc.contributor.authorVenkitaraman, Ashoken
dc.contributor.authorSkylaris, Chris-Kritonen
dc.date.accessioned2017-08-08T11:45:25Z
dc.date.available2017-08-08T11:45:25Z
dc.date.issued2011-07en
dc.identifier.issn1553-734X
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/266005
dc.description.abstractThe breast cancer suppressor BRCA2 controls the recombinase RAD51 in the reactions that mediate homologous DNA recombination, an essential cellular process required for the error-free repair of DNA double-stranded breaks. The primary mode of interaction between BRCA2 and RAD51 is through the BRC repeats, which are ∼35 residue peptide motifs that interact directly with RAD51 in vitro. Human BRCA2, like its mammalian orthologues, contains 8 BRC repeats whose sequence and spacing are evolutionarily conserved. Despite their sequence conservation, there is evidence that the different human BRC repeats have distinct capacities to bind RAD51. A previously published crystal structure reports the structural basis of the interaction between human BRC4 and the catalytic core domain of RAD51. However, no structural information is available regarding the binding of the remaining seven BRC repeats to RAD51, nor is it known why the BRC repeats show marked variation in binding affinity to RAD51 despite only subtle sequence variation. To address these issues, we have performed fluorescence polarisation assays to indirectly measure relative binding affinity, and applied computational simulations to interrogate the behaviour of the eight human BRC-RAD51 complexes, as well as a suite of BRC cancer-associated mutations. Our computational approaches encompass a range of techniques designed to link sequence variation with binding free energy. They include MM-PBSA and thermodynamic integration, which are based on classical force fields, and a recently developed approach to computing binding free energies from large-scale quantum mechanical first principles calculations with the linear-scaling density functional code onetep. Our findings not only reveal how sequence variation in the BRC repeats directly affects affinity with RAD51 and provide significant new insights into the control of RAD51 by human BRCA2, but also exemplify a palette of computational and experimental tools for the analysis of protein-protein interactions for chemical biology and molecular therapeutics.
dc.languageengen
dc.publisherPublic Library of Science (PLoS)
dc.rightsAttribution 4.0 Internationalen
dc.rightsAttribution 4.0 Internationalen
dc.rightsAttribution 4.0 Internationalen
dc.rightsAttribution 4.0 Internationalen
dc.rightsAttribution 4.0 Internationalen
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en
dc.subjectAmino Acid Motifsen
dc.subjectAmino Acid Sequenceen
dc.subjectBRCA2 Proteinen
dc.subjectFluorescence Polarization Immunoassayen
dc.subjectHumansen
dc.subjectModels, Molecularen
dc.subjectMolecular Sequence Dataen
dc.subjectMutationen
dc.subjectProtein Bindingen
dc.subjectProtein Interaction Domains and Motifsen
dc.subjectProtein Interaction Mappingen
dc.subjectProtein Stabilityen
dc.subjectRad51 Recombinaseen
dc.subjectRepetitive Sequences, Amino Aciden
dc.subjectSequence Alignmenten
dc.subjectThermodynamicsen
dc.titleInterrogation of the protein-protein interactions between human BRCA2 BRC repeats and RAD51 reveals atomistic determinants of affinity.en
dc.typeArticle
prism.endingPagee1002096
prism.issueIdentifier7en
prism.publicationDate2011en
prism.publicationNamePLoS Computational Biologyen
prism.startingPagee1002096
prism.volume7en
dc.identifier.doi10.17863/CAM.9746
dcterms.dateAccepted2011-05-04en
rioxxterms.versionofrecord10.1371/journal.pcbi.1002096en
rioxxterms.licenseref.urihttp://creativecommons.org/licenses/by/4.0/en
rioxxterms.licenseref.startdate2011-07en
dc.contributor.orcidPayne, Michael [0000-0002-5250-8549]
dc.identifier.eissn1553-7358
rioxxterms.typeJournal Article/Reviewen
pubs.funder-project-idMRC (G0600332)
pubs.funder-project-idMRC (G0700651)
pubs.funder-project-idMRC (G9900064)
pubs.funder-project-idEPSRC (EP/F032773/1)
cam.issuedOnline2011-07-14en
dc.identifier.urlhttps://journals.plos.org/ploscompbiol/article?id=10.1371/journal.pcbi.1002096en
cam.orpheus.successThu Jan 30 12:59:23 GMT 2020 - The item has an open VoR version.*
rioxxterms.freetoread.startdate2100-01-01


Files in this item

Thumbnail
Thumbnail

This item appears in the following Collection(s)

Show simple item record

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