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1、TheEMBOJournalVol.16No.18pp.5730±5741,1997DualroleoftheC34subunitofRNApolymeraseIIIintranscriptioninitiationIsabelleBrun,AndreÂSentenacandTFIIIBisthegeneraltranscriptionfactorrecognizedbyMichelWerner1polIIIsinceaTFIIIB´DNAcomplexcandirectmultipleroundsoftranscrip
2、tioninvitro(Kassavetisetal.,1990).ServicedeBiochimieetGeÂneÂtiqueMoleÂculaire,BaÃtiment142,TFIIIBiscomposedofthreepolypeptides,TATA-bindingCEA/Saclay,F-91191Gif-sur-YvetteCedex,Franceprotein(TBP;HuetandSentenac,1992;Kassavetisetal.,1Correspondingauthor1992b),agen
3、eraltranscriptionfactorrequiredfortran-e-mail:werner@jonas.saclay.cea.frscriptionbyalleukaryoticandarchaebacterialRNApoly-merases(Struhl,1995,andreferencestherein),TFIIIB90,TheC34subunitofyeastRNApolymerase(pol)IIIisa90kDasubunitwhichhasnoequivalentamongtheotherp
4、artofasubcomplexofthreesubunitswhichhavenoRNApolymerasegeneraltranscriptionfactors(KassavetiscounterpartintheothertwonuclearRNApolymerases.etal.,1995;Robertsetal.,1996;RuÈthetal.,1996),andThissubunitinteractswithTFIIIB70andisthereforeTFIIIB70,a70kDaproteinwhichis
5、homologoustothoughttoparticipateinpolIIIrecruitment.TostudyarchaealgeneralfactorTFB(HausnerandThomm,1995;theroleofC34intranscription,wehavemutagenizedQureshietal.,1995)andtopolIIfactorTFIIB(BuratowskiRPC34,thegeneencodingC34,andfoundthatandZhou,1992;ColbertandHah
6、n,1992;LoÂpez-De-LeoÂnmutationsaffectinggrowthalsoalteredC34interactionetal.,1992).TFIIBisthelastgeneraltranscriptionfactorwithTFIIIB70.ThetwomutantpolIIIthatweretoentertheclassIIpre-initiationcomplexbeforepolIIpuri®edhadcatalyticpropertiesindistinguishablefrom(B
7、uratowskietal.,1989),pointingtothepossibilitythatthoseofthewild-typepolIIIonapoly[d(A±T)]tem-TFIIIB70mightsimilarlyrecruitpolIII.plate,whilespeci®ctranscriptionofpolIIIgenesThreepolIIIsubunits,C82,C34andC31,thathaveinthepresenceofgeneraltranscriptionfactorswasnoc
8、ounterpartintheotherRNApolymerases(Mosrinimpaired.ThedefectoftheC34-1124mutantenzymeetal.,1990;Chiannilkulchaietal.,1992;Stettleretal.,couldbecompensatedbyincr
