Comparative Genomics Identies the Mouse Bmp3 Promoter and an Upstream Evolutionary Conserved Region ECR) in Mammals.docVIP

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Comparative Genomics Identies the Mouse Bmp3 Promoter and an Upstream Evolutionary Conserved Region ECR) in Mammals.doc

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Comparative Genomics Identies the Mouse Bmp3 Promoter and an Upstream Evolutionary Conserved Region ECR) in Mammals

ComparativeGenomicsIdentifiestheMouseBmp3 PromoterandanUpstreamEvolutionaryConserved Region(ECR)inMammals JonathanW.Lowery,AnnaW.LaVigne,ShoichiroKokabu,VickiRosen* DepartmentofDevelopmentalBiology,HarvardSchoolofDentalMedicine,Boston,Massachusetts,UnitedStatesofAmerica Abstract TheBoneMorphogeneticProtein(BMP)pathwayisamulti-membersignalingcascadewhosebasiccomponentsarefound inallanimals.Onemember,BMP3,whicharosemorerecentlyinevolutionandisfoundonlyindeuterostomes,servesa uniqueroleasanantagonisttoboththecanonicalBMPandActivinpathways.However,themechanismsthatcontrolBMP3 expression,andthecis-regulatoryregionsmediatingthisregulation,remainpoorlydefined.Withthisinmind,wesoughtto identifytheBmp3promoterinmouse(M.musculus)throughfunctionalandcomparativegenomicanalyses.Wefoundthat theminimalpromoterrequiredforexpressioninresideswithin0.8kbupstreamofBmp3inaregionthatishighlyconserved withrat(R.norvegicus).Wealsofoundthatanupstreamregionabuttingtheminimalpromoteractsasarepressorofthe minimal promoter in HEK293T cells and osteoblasts. Strikingly, a portion of this region is conserved among all available eutherianmammalgenomes(47/47),butnotinanynon-eutheriananimal(0/136).Wealsoidentifiedmultipleconserved transcription factor binding sites in the Bmp3 upstream ECR, suggesting that this region may preserve common cis- regulatoryelementsthatgovernBmp3expressionacrosseutherianmammals.SincedysregulationofBMPsignalingappears toplayaroleinhumanhealthanddisease,ourfindingsmayhaveapplicationinthedevelopmentofnoveltherapeutics aimedatmodulatingBMPsignalinginhumans. Citation: Lowery JW, LaVigne AW, Kokabu S, Rosen V (2013) Comparative Genomics Identifies the Mouse Bmp3 Promoter and an Upstream Evolutionary ConservedRegion(ECR)inMammals.PLoSONE8(2):e57840.doi:10.1371/journal.pone.0057840 Editor:BerndSokolowski,UniversityofSouthFlorida,UnitedStatesofAmerica ReceivedNovember6,2012;AcceptedJanuary26,2013;PublishedFebruary22,2013 Copyright: ? 2013 Lowery et al. This is an open-access art