a)Corepromoter-35 Box;-35site:RNApol.looselybinding siteRNApol. recognition site (R site)TTGAC(Sextama Box)-10 site: RNApol.firmly binding site-10Box/PribnowBox;RNApol. Binding template strand (B site)TATAAT (pribnow-schaller Box 1975)-35 (R)-10 (B)
a）Core promoter -35 Box ; RNApol. recognition site (R site) TTGAC (Sextama Box) -35 site: RNApol. loosely binding site -10 Box/Pribnow Box ; TATAAT (pribnow-schaller Box 1975) -10 site: RNApol. firmly binding site RNApol. Binding template strand (B site) -35 (R) -10 (B)
PNAS 1975.Feb.V.72 N.2Schaller Heidberd U Ger.Proc.Nat.AoadSei.USAHeinzVol. 72, No. 2, pp.737-741, February 1975Nucleotide Sequence of an RNAPolymerase Binding Site from the DNABacteriophage fdof(promoters/DNAsequencing/protein-DNA interaction)HEINZSCHALLER,CHRISTOPHERGRAY,AND KARINHERRMANNMax-Planck-lastitus furVirusforscbung.Tebingen and Lehrstuhl fur Mikrobiologie derUniversitss Heidelberg, 6goo HeidelbergImNeuenheimerYeld280,WestGermanCommunicoted by H, Gobind Khorana, DecemberABSTRACTThe primarystructure ofPNAS 1975.Mar.V.72 N.3Proe.Naf.Acad.Sei., USApolymerase binding bite l the replientiveVol. 72, No, 3, pp. 784-788, March 1975phage fd has been determined by direct DNItistDavid Pribnow HarvardUT.G-C-T-T-C-T-G-A-C-T-A-T-A-A-T-A-C-A-C.A-C-G-A-A-G-A-C-T-G-A-T-A-T-T-A-T.C-TG-A-G-A-C-C-T-G-A-T-T-T-T-T-G-A3NucleotideSequence of an RNAPolymeraseBinding Siteat anT-C-T-G-G-A-C-T-A-A-1T7oleelregions with2-faldPromoterEarly(initlated complex/DNA-RNAhybridization/RNAsequence)terofthehindinDAVID PRIBNOWPromoters contain, as an essential part,a sitefand stable attachment of the enayme RNA poThe Biological Laboratories, Harvard University, Cambridge, Masachusette oziasensuringe strand- and aite-specifie initiationCommunicatedby J.D,Watson,December1e,1974poymerasebinding sitescan beishainHABSTRACTEscherichiacoliRNA(ECTight complexes between RNA polymerase and its bindingpol2.7.7.6), bandinatightcomplexcarlyTT promoter,sites are destabilised when the salt concentration in the bind.-ofDNAfromto43basdigestionbproteetsing mixture is raised (or when the temperature is lowered)DNase I.Theeeted DNA fragmentcontainsboth the(2-4, 6, 21-23). Under these eonditions DNasse can degradesiteforRNApolyrerase and themRNAinitiationbindall of the DNA. If, however, the polymerase has been allowedof theDNApromoter.mRNAtto initiateRNA synthesis,elevated salt concentrations do notdisengage the polymerase (2, 21-23), and polymerase-pro-tected DNA fragments ean be obtained (13).Ihavebeenable to selectively"lock"RNApolymerase ontotheT7 A3The transcription of a gene begins at a specifie site called apromoter by forming a salt-stable, initiated complexex?promoter(1),At such a sitetheRNApolymerase interactsclusively at that site on the T7 genome. The key to thewith the DNA in order to initiate the syntheais of an RNAspecification of the A3 site is the use of a dinucleotide (24, 25)moleeule.HowdoesthepolymeraseTecoonepromoter.to direet a limited initiation event. E, Minkley and I (20)
PNAS 1975.Feb. V.72 N.2 Heinz Schaller Heidberd U Ger. PNAS 1975.Mar.V.72 N.3 David Pribnow Harvard U
a)Corepromoter-35 Box ;-35site RNApol.loosely bindingsiteRNApol. recognition site (R site)-10 site RNApol.firmly binding sitePribnowBox;RNApol. Binding template strand (B site)Initiation site;+1 RNAtranscriptional startpoint (Isite)A/G-35 (R)-10 (B)+1 ()RNA
a）Core promoter -35 Box ; RNApol. recognition site (R site) -35 site RNApol. loosely binding site Pribnow Box ; -10 site RNApol. firmly binding site RNApol. Binding template strand (B site) Initiation site ; +1 RNA transcriptional startpoint (I site) A/G +1 (I) RNA -35 (R) -10 (B)
b)UpelementCAP-cAMPbindingsite(ActivatorregionAR)IRIR-50-40-70ARIARIIAR I;其中的IR是CAP-cAMP的强结合位点（-70~-50）CAP-cAMP的弱结合位点（-50～-40）AR Il;ARI+CAP-cAMP提高ARII的结合效率cooperative effect
b) Up element CAP-cAMP binding site (Activator region AR) ● ARI + CAP-cAMP ● AR I; 其中的IR是CAP-cAMP 的强结合位点（-70 ~ -50） ● AR II; CAP-cAMP 的弱结合位点（-50 ~ -40） cooperative effect 提高ARII 的结合效率 IR -70 AR I -40 AR II IR -50
ARII+CAP-CAMPpromotionRNApol. Into-35BoxintoPribnowBoxstarting transcriptionAR+CAP-CAMP复合体促使-35Box附近GC岛区的双螺旋结构稳定性降低PribnowBox的解链温度降低，利于转录启动RNApolARIARIIGC Island-35boxPribnowbox
RNApol AR II + CAP-cAMP 复合体 促使-35 Box 附近GC岛区的双螺旋结构稳定性降低， Pribnow Box 的解链温度降低，利于转录启动 ARI ARII GC Island -35box Pribnow box ● AR II + CAP-cAMP promotion RNApol. Into -35 Box into Pribnow Box starting transcription RNApol RNApol