Nucleolin Participates in DNA Double-Strand Break-Induced Damage Response through MDC1-Dependent Pathway 英文参考文献.docVIP

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Nucleolin Participates in DNA Double-Strand Break-Induced Damage Response through MDC1-Dependent Pathway 英文参考文献.doc

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Nucleolin Participates in DNA Double-Strand Break-Induced Damage Response through MDC1-Dependent Pathway 英文参考文献

NucleolinParticipatesinDNADouble-StrandBreak- InducedDamageResponsethroughMDC1-Dependent Pathway JunyaKobayashi1*,HirokoFujimoto1,JunSato1,IkueHayashi2,SandeepBurma3,ShinyaMatsuura4, DavidJ.Chen3,KenshiKomatsu1 1Department ofGenomeRepairDynamics,RadiationBiologyCenter,KyotoUniversity, Kyoto,Japan, 2GraduateSchoolofBiomedicalSciences,HiroshimaUniversity, Hiroshima,Japan,3DivisionofMolecularRadiationBiology,DepartmentofRadiationOncology,UniversityofTexasSouthwesternMedicalCenteratDallas,Dallas,Texas, UnitedStatesofAmerica,4DepartmentofGeneticsandCellBiology,ResearchInstituteforRadiationBiologyandMedicine,HiroshimaUniversity,Hiroshima,Japan Abstract H2AXisanimportantfactorforchromatinremodelingtofacilitateaccumulationofDNAdamage-relatedproteinsatDNA double-strandbreak(DSB)sites.InordertofurtherunderstandtheroleofH2AXintheDNAdamageresponse(DDR),we attempted to identify H2AX-interacting proteins by proteomics analysis. As a result, we identified nucleolin as one of candidates.Here,weshowanovelroleofamajornucleolarprotein,nucleolin,inDDR.Nucleolininteractedwithc-H2AX and accumulated to laser micro-irradiated DSB damage sites. Chromatin Immunoprecipitation assay also displayed the accumulation of nucleolin around DSB sites. Nucleolin-depleted cells exhibited repression of both ATM-dependent phosphorylation following exposure to c-ray and subsequent cell cycle checkpoint activation. Furthermore, nucleolin- knockdownreducedHRandNHEJactivityandshoweddecreaseinIR-inducedchromatinaccumulationofHR/NHEJfactors, agreeingwiththedelayedkineticsofc-H2AXfocus.Moreover,nucleolin-knockdowndecreasedMDC1-relatedeventssuch as focus formation of 53BP1, RNF168, phosphorylated ATM, and H2A ubiquitination. Nucleolin also showed FACT-like activity for DSB damage-induced histone eviction from chromatin. Taken together, nucleolin could promote both ATM- dependentcellcyclecheckpointandDSBrepairbyfunctioninginanMDC1-relatedpathwaythroughitsFACT-likefunction. Citation:KobayashiJ,FujimotoH,Sato