Cell Cycle-Dependent Induction of Homologous Recombination by a Tightly Regulated I-SceI Fusion Protein 英文参考文献.docVIP

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Cell Cycle-Dependent Induction of Homologous Recombination by a Tightly Regulated I-SceI Fusion Protein 英文参考文献.doc

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Cell Cycle-Dependent Induction of Homologous Recombination by a Tightly Regulated I-SceI Fusion Protein 英文参考文献

CellCycle-DependentInductionofHomologous RecombinationbyaTightlyRegulatedI-SceIFusion Protein AndreaHartlerode,ShobuOdate¤a,InboShim¤b,JeniferBrown¤c,RalphScully* DepartmentofMedicine,HarvardMedicalSchoolandBethIsraelDeaconessMedicalCenter,Boston,Massachusetts,UnitedStatesofAmerica Abstract Double-strand break repair is executed by two major repair pathways: non-homologous end joining (NHEJ) and homologousrecombination(HR).WhereasNHEJcontributestotherepairofionizingradiation(IR)-induceddoublestrand breaks(DSBs)throughoutthecellcycle,HRactspredominantlyduringtheSandG2phasesofthecellcycle.Therare-cutting restrictionendonuclease,I-SceI,isincommonusetostudytherepairofsite-specificchromosomalDSBsinvertebratecells. TofacilitateanalysisofI-SceI-inducedDSBrepair,wehavedevelopedastablyexpressedI-SceIfusionproteinthatenables precise temporal control of I-SceI activation, and correspondingly tight control of the timing of onset of site-specific chromosome breakage. I-SceI-induced HR showed a strong, positive linear correlation with the percentage of cells in S phase,andwasnegativelycorrelatedwiththeG1fraction.AcutedepletionofBRCA1,akeyregulatorofHR,disruptedthe relationship between S phase fraction and I-SceI-induced HR, consistent with the hypothesis that BRCA1 regulates HR duringSphase. Citation:HartlerodeA,OdateS,ShimI,BrownJ,ScullyR(2011)CellCycle-DependentInductionofHomologousRecombinationbyaTightlyRegulatedI-SceI FusionProtein.PLoSONE6(3):e16501.doi:10.1371/journal.pone.0016501 Editor:WafikEl-Deiry,UniversityofPennsylvania,UnitedStatesofAmerica ReceivedDecember7,2010;AcceptedDecember30,2010;PublishedMarch9,2011 Copyright: ?2011 Hartlerodeet al. This is an open-access articledistributed under the terms of the Creative Commons Attribution License, which permits unrestricteduse,distribution,andreproductioninanymedium,providedtheoriginalauthorandsourcearecredited. Funding: NIH R01CA095175; R01GM073894; R21CA144022. The funders had no role in study design, data collection a