DNA Polymerase α (swi7) and the Flap Endonuclease Fen1 (rad2) Act Together in the S-Phase Alkylation Damage Response in S. pombe 英文参考文献.docVIP

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DNA Polymerase α (swi7) and the Flap Endonuclease Fen1 (rad2) Act Together in the S-Phase Alkylation Damage Response in S. pombe 英文参考文献.doc

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DNA Polymerase α (swi7) and the Flap Endonuclease Fen1 (rad2) Act Together in the S-Phase Alkylation Damage Response in S. pombe 英文参考文献

DNAPolymerasea(swi7)andtheFlapEndonuclease Fen1(rad2)ActTogetherintheS-PhaseAlkylation DamageResponseinS.pombe MilanaKoulintchenko1,SonyaVengrova1,TrevorEydmann1,PrakashArumugam2,JacobZ.Dalgaard1* 1DivisionofBiomedicalCellBiology,WarwickMedicalSchool,GibbetHillCampus,UniversityofWarwick,Coventry,UnitedKingdom,2SchoolofLifeSciences,GibbetHill Campus,UniversityofWarwick,Coventry,UnitedKingdom Abstract Polymerase a is an essential enzyme mainly mediating Okazaki fragment synthesis during lagging strand replication. A specific point mutation in Schizosaccharomyces pombe polymerase a named swi7-1, abolishes imprinting required for mating-type switching. Here weinvestigatewhether this mutationconfers anygenome-wide defects.We showthat the swi7-1 mutation renders cells hypersensitive to the DNA damaging agents methyl methansulfonate (MMS), hydroxyurea (HU)andUVandincapacitatesactivationoftheintra-ScheckpointinresponsetoDNAdamage.Inadditionweshowthat,in theswi7-1background,cellsarecharacterizedbyanelevatedlevelofrepairfociandrecombination,indicativeofincreased geneticinstability.Furthermore,wedetectnovelSwi1-,-Swi3-andPola-dependentalkylationdamagerepairintermediates withmobilityon2D-gelthatsuggestspresenceofsingle-strandedregions.Geneticinteractionstudiesshowedthattheflap endonucleaseFen1worksinthesamepathwayasPolaintermsofalkylationdamageresponse.Fen1wasalsorequiredfor formationofalkylation-damagespecificrepairintermediates.WeproposeamodeltoexplainhowPola,Swi1,Swi3and Fen1mightacttogethertodetectandrepairalkylationdamageduringS-phase. Citation: Koulintchenko M, Vengrova S, Eydmann T, Arumugam P, Dalgaard JZ (2012) DNA Polymerase a (swi7) and the Flap Endonuclease Fen1 (rad2 ) Act TogetherintheS-PhaseAlkylationDamageResponseinS.pombe.PLoSONE7(10):e47091.doi:10.1371/journal.pone.0047091 Editor:MartinG.Marinus,UniversityofMassachusettsMedicalSchool,UnitedStatesofAmerica ReceivedJuly27,2012;AcceptedSeptember7,2012;PublishedOctober11,2012 Copyright:?2012Koulintchenkoetal.Thi