Expression of 5 S rRNA genes linked to 35 S rDNA in plants, their epigenetic modification and regulatory element divergence.docVIP

Expression of 5 S rRNA genes linked to 35 S rDNA in plants, their epigenetic modification and regulatory element divergence.doc

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Expression of 5 S rRNA genes linked to 35 S rDNA in plants, their epigenetic modification and regulatory element divergence

Garciaetal.BMCPlantBiology2012,12:95 /1471-2229/12/95 RESEARCH ARTICLE OpenAccess Expressionof5SrRNAgeneslinkedto35SrDNA inplants,theirepigeneticmodificationand regulatoryelementdivergence SòniaGarcia1,LucieCrhákKhaitová2andAle?Kova?ík2* Abstract Background:Inplants,the5SrRNAgenesusuallyoccurasseparatetandems(S-typearrangement)or,less commonly,linkedto35SrDNAunits(L-type).Theactivityoflinkedgenesremainsunknownsofar.Westudiedthe homogeneityandexpressionof5SgenesinseveralspeciesfromfamilyAsteraceaeknowntocontainlinked35S- 5Sunits.Additionally,theirmethylationstatuswasdeterminedusingbisulfitesequencing.Fluorescenceinsitu hybridizationwasappliedtorevealthesub-nuclearpositionsofrDNAarrays. Results:WefoundthathomogenizationofL-typeunitswenttocompletioninmost(4/6)butnotallspecies.Two speciescontainedmajorL-typeandminorS-typeunits(termedLs-type).Thelinkedgenesdominate5SrDNA expressionwhiletheseparatetandemsdonotseemtobeexpressed.MembersoftribeAnthemideaeevolved functionalvariantsofthepolymeraseIIIpromoterinwhicharesidingC-boxelementdiffersfromthecanonical angiospermmotifbyasmuchas30%.Onthisbasis,amorerelaxedconsensussequenceofaplantC-box:(5’- RGSWTGGGTG-3’)isproposed.The5SparalogsdisplayheavyDNAmethylationsimilarlyastotheirunlinked counterparts.FISHrevealedthecloseassociationof35S-5Sarrayswithnucleolarperipheryindicatingthat transcriptionof5Sgenesmayoccurinthisterritory. Conclusions:Weshowthattheunusuallinkedarrangementof5Sgenes,occurringinseveralplantspecies,isfully compatiblewiththeirexpressionandfunctionality.Thisextraordinary5Sgenedynamicsismanifestedatdifferent levels,suchasvariationinintrachromosomalpositions,unitstructure,epigeneticmodificationandconsiderable divergenceofregulatorymotifs. Keywords:5Sexpression,Artemisia,Asteraceae,C-box,DNAmethylation,PolIIIpromoterdivergence,rDNA organization Background 18S,5.8Sand26SrRNAgenes,theinternaltranscribed NuclearribosomalDNA(rDNA)encoding5S,5.8S,18S spacers(ITSs),andanintergenicspacer(IGS)(forreview and26SrRNAbelon

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