Biopolym. Cell. 1988; 4(1):7-14.
Structure and Function of Biopolymers
The interaction of Escherichia coli RNA polymerase with DNA-duplexes containing repeats. The influence of the nucleotides' sequence
1Koroleva O. N., 1Drutsa V. L., 1Shabarova Z. A.
  1. M. V. Lomonosov State University
    Moscow, USSR


The chemical-enzymatic synthesis of DNA-duplexes d(pCATTATAATG)n (I), d(pGATTATAATC)n (II), d(pCTTTATAAAG)n (III), d(pCATTTAAATG)n (IV), d(pCAATATATTG)n (V), d(pCATAATTATG)n (VI) (n= 104-20), containing repeats of the sequences completely and partially homologous to the «ideal» Pribnow box (TATAATG) has been carried out. The binding of E. coli RNA polymerase to synthetic DNA duplexes and in vitro transcription of these duplexes have been studied. It has been shown that duplexes I, II and V containing the sequence TATA-T form heparin-resistant Uanscriptionally active complexes with enzyme (the half life is ~ 100-200 min) while duplexes III and IV do not form stable complexes. The pnedominant in vitro transcription initiation sites were revealed within duplexes I, II and VI.


[1] Koroleva ON, Drutsa VL, Dolinnaia NG, Tsytovich AV, Shabarova ZA. DNA-like duplexes containing repetitive sequences. VII. Chemico-enzymatic synthesis of polymers with fragments of natural promotors. Mol Biol (Mosk). 1984;18(1):146-60.
[2] Koroleva ON, Shabarova ZA, Bibilashvili RSh. Interaction of Escherichia coli RNA-polymerase with DNA-duplexes containing repetitive sequences. Mol Biol (Mosk). 1985;19(2):516-23.
[3] Koroleva ON, Drutsa VL, Shabarova ZA. Transcription of the synthetic DNA duplexes with repeats of promoter fragments. Biopolym. Cell. 1985; 1(6):307-11.
[4] Matteucci MD, Caruthers MH. Synthesis of deoxyoligonucleotides on a polymer support. J Amer Chem Soc. 1981; 103(11):3185-3191.
[5] Maxam AM, Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499-560.
[6] Strauss HS, Boston RS, Record MT Jr, Burgess RR. Variables affecting the selectivity and efficiency of retention of DNA fragments by E. coli RNA polymerase in the nitrocellulose-filter-binding assay. Gene. 1981;13(1):75-87.
[7] Downey KM, Jurmark BS, So AG. Determination of nucleotide sequences at promoter regions by the use of dinucleotides. Biochemistry. 1971;10(26):4970-5.
[8] Siebenlist U, Simpson RB, Gilbert W. E. coli RNA polymerase interacts homologously with two different promoters. Cell. 1980;20(2):269-81.
[9] von Hippel PH, Bear DG, Winter RB, Berg OG. Molecular aspects of promoter function: an overview. In: Promoters: structure and function . Eds R. L. Rodriguez, M. J. Chamberlin. New York: Praeger Publ., 1982:3-34.
[10] Dairi T, Inokuchi K, Mizuno T, Mizushima S. Positive control of transcription initiation in Escherichia coli. A base substitution at the Pribnow box renders ompF expression independent of a positive regulator. J Mol Biol. 1985;184(1):1-6.