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dc.contributor.authorDavies, Owen Ren
dc.contributor.authorForment, Josepen
dc.contributor.authorSun, Meidaien
dc.contributor.authorBelotserkovskaya, Rimmaen
dc.contributor.authorCoates, Juliaen
dc.contributor.authorGalanty, Yaronen
dc.contributor.authorDemir, Mukerremen
dc.contributor.authorMorton, Christopheren
dc.contributor.authorRzechorzek, Neilen
dc.contributor.authorJackson, Stephenen
dc.contributor.authorPellegrini, Lucaen
dc.date.accessioned2015-03-09T11:56:04Z
dc.date.available2015-03-09T11:56:04Z
dc.date.issued2015-01-05en
dc.identifier.citationNature Structural & Molecular Biology, 22, 150–157 (2015) doi: 10.1038/nsmb.2937en
dc.identifier.issn1545-9993
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/246946
dc.description.abstractMammalian CtIP protein has major roles in DNA double-strand break (DSB) repair. Although it is well established that CtIP promotes DNA-end resection in preparation for homology-dependent DSB repair, the molecular basis for this function has remained unknown. Here we show by biophysical and X-ray crystallographic analyses that the N-terminal domain of human CtIP exists as a stable homotetramer. Tetramerization results from interlocking interactions between the N-terminal extensions of CtIP’s coiled-coil region, which lead to a ‘dimer-of-dimers’ architecture. Through interrogation of the CtIP structure, we identify a point mutation that abolishes tetramerization of the N-terminal domain while preserving dimerization in vitro. Notably, we establish that this mutation abrogates CtIP oligomer assembly in cells, thus leading to strong defects in DNA-end resection and gene conversion. These findings indicate that the CtIP tetramer architecture described here is essential for effective DSB repair by homologous recombination.
dc.description.sponsorshipWe thank M. Kilkenny for help with the collection of X-ray diffraction data, A. Sharff and P. Keller for help with X-ray data processing and J.D. Maman for assistance with SEC-MALS. This work was supported by a Wellcome Trust Senior Research Fellowship award in basic biomedical sciences (L.P.), an Isaac Newton Trust research grant (L.P. and O.R.D.) and a Cambridge Overseas Trust PhD studentship (M.D.S.). Research in the laboratory of S.P.J. is funded by Cancer Research UK (CRUK; programme grant C6/A11224), the European Research Council and the European Community Seventh Framework Programme (grant agreement no. HEALTH-F2-2010-259893 (DDResponse)). Core funding is provided by Cancer Research UK (C6946/A14492) and the Wellcome Trust (WT092096). S.P.J. receives his salary from the University of Cambridge, supplemented by CRUK. J.V.F. is funded by Cancer Research UK programme grant C6/A11224 and the Ataxia Telangiectasia Society. R.B. and J.C. are funded by Cancer Research UK programme grant C6/A11224. Y.G. and M.D. are funded by the European Research Council grant DDREAM.
dc.languageEnglishen
dc.language.isoenen
dc.publisherNature Publishing Group
dc.titleCtIP tetramer assembly is required for DNA-end resection and repairen
dc.typeArticle
dc.description.versionThis is the accepted manuscript of a paper published in Nature Structural & Molecular Biology, 22, 150–157 (2015) doi: 10.1038/nsmb.2937en
prism.endingPage157
prism.publicationDate2015en
prism.publicationNameNature Structural & Molecular Biologyen
prism.startingPage150
dc.rioxxterms.funderCRUK
dc.rioxxterms.funderWellcome Trust
dc.rioxxterms.funderERC
dc.rioxxterms.projectidC6946/A14492
dc.rioxxterms.projectidWT092096
dcterms.dateAccepted2014-11-21en
rioxxterms.versionofrecord10.1038/nsmb.2937en
rioxxterms.licenseref.urihttp://www.rioxx.net/licenses/all-rights-reserveden
rioxxterms.licenseref.startdate2015-01-05en
dc.contributor.orcidForment, Josep [0000-0002-7797-2583]
dc.contributor.orcidGalanty, Yaron [0000-0001-7167-9004]
dc.contributor.orcidJackson, Stephen [0000-0001-9317-7937]
dc.contributor.orcidPellegrini, Luca [0000-0002-9300-497X]
dc.identifier.eissn1545-9985
rioxxterms.typeJournal Article/Reviewen
pubs.funder-project-idCancer Research UK (11224)
pubs.funder-project-idWELLCOME TRUST (104641/Z/14/Z)
pubs.funder-project-idEuropean Research Council (268536)
pubs.funder-project-idWellcome Trust (084279/Z/07/A)
pubs.funder-project-idWellcome Trust (092096/Z/10/Z)
pubs.funder-project-idCancer Research UK (A14492)
rioxxterms.freetoread.startdate2015-07-05


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