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. 1989 May 25;17(10):3855–3863. doi: 10.1093/nar/17.10.3855

Effect of the sequence-dependent structure of the 17 bp AT spacer on the strength of consensuslike E.coli promoters in vivo.

T Lozinski 1, W T Markiewicz 1, T K Wyrzykiewicz 1, K L Wierzchowski 1
PMCID: PMC317864  PMID: 2660103

Abstract

Three E.coli promoters with the consensus sequences in the -35 and -10 regions and the 17 bp spacer made of random, heteronomous, and of both these classes of AT DNA simultaneously were constructed and cloned into plasmid pDS3. Electrophoretic gel mobilities of restriction fragments containing these promoters indicated that bending of the latter was proportional to the number of heteronomous AT DNA tracts. The strength of these promoters in vivo measured in relation to an internal transcriptional standard was shown to correlate well with gel mobilities of the respective restriction fragments and to decrease with the number of potential DNA bending sites encoded in the promoter structure.

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Selected References

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  1. Aoyama T., Takanami M., Ohtsuka E., Taniyama Y., Marumoto R., Sato H., Ikehara M. Essential structure of E. coli promoter: effect of spacer length between the two consensus sequences on promoter function. Nucleic Acids Res. 1983 Sep 10;11(17):5855–5864. doi: 10.1093/nar/11.17.5855. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Auble D. T., Allen T. L., deHaseth P. L. Promoter recognition by Escherichia coli RNA polymerase. Effects of substitutions in the spacer DNA separating the -10 and -35 regions. J Biol Chem. 1986 Aug 25;261(24):11202–11206. [PubMed] [Google Scholar]
  3. Borowiec J. A., Gralla J. D. All three elements of the lac ps promoter mediate its transcriptional response to DNA supercoiling. J Mol Biol. 1987 May 5;195(1):89–97. doi: 10.1016/0022-2836(87)90329-9. [DOI] [PubMed] [Google Scholar]
  4. Borowiec J. A., Gralla J. D. Supercoiling response of the lac ps promoter in vitro. J Mol Biol. 1985 Aug 20;184(4):587–598. doi: 10.1016/0022-2836(85)90305-5. [DOI] [PubMed] [Google Scholar]
  5. Buc H., McClure W. R. Kinetics of open complex formation between Escherichia coli RNA polymerase and the lac UV5 promoter. Evidence for a sequential mechanism involving three steps. Biochemistry. 1985 May 21;24(11):2712–2723. doi: 10.1021/bi00332a018. [DOI] [PubMed] [Google Scholar]
  6. Burgess R. R., Jendrisak J. J. A procedure for the rapid, large-scall purification of Escherichia coli DNA-dependent RNA polymerase involving Polymin P precipitation and DNA-cellulose chromatography. Biochemistry. 1975 Oct 21;14(21):4634–4638. doi: 10.1021/bi00692a011. [DOI] [PubMed] [Google Scholar]
  7. Deuschle U., Kammerer W., Gentz R., Bujard H. Promoters of Escherichia coli: a hierarchy of in vivo strength indicates alternate structures. EMBO J. 1986 Nov;5(11):2987–2994. doi: 10.1002/j.1460-2075.1986.tb04596.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Drew H. R., Weeks J. R., Travers A. A. Negative supercoiling induces spontaneous unwinding of a bacterial promoter. EMBO J. 1985 Apr;4(4):1025–1032. doi: 10.1002/j.1460-2075.1985.tb03734.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ehrlich R., Larousse A., Jacquet M. A., Marin M., Reiss C. In vitro transcription initiation from three different Escherichia coli promoters. Effect of supercoiling. Eur J Biochem. 1985 Apr 15;148(2):293–298. doi: 10.1111/j.1432-1033.1985.tb08838.x. [DOI] [PubMed] [Google Scholar]
  10. Gamper H. B., Hearst J. E. A topological model for transcription based on unwinding angle analysis of E. coli RNA polymerase binary, initiation and ternary complexes. Cell. 1982 May;29(1):81–90. doi: 10.1016/0092-8674(82)90092-7. [DOI] [PubMed] [Google Scholar]
  11. Hawley D. K., McClure W. R. Compilation and analysis of Escherichia coli promoter DNA sequences. Nucleic Acids Res. 1983 Apr 25;11(8):2237–2255. doi: 10.1093/nar/11.8.2237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kirkegaard K., Buc H., Spassky A., Wang J. C. Mapping of single-stranded regions in duplex DNA at the sequence level: single-strand-specific cytosine methylation in RNA polymerase-promoter complexes. Proc Natl Acad Sci U S A. 1983 May;80(9):2544–2548. doi: 10.1073/pnas.80.9.2544. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Koo H. S., Wu H. M., Crothers D. M. DNA bending at adenine . thymine tracts. Nature. 1986 Apr 10;320(6062):501–506. doi: 10.1038/320501a0. [DOI] [PubMed] [Google Scholar]
  14. Menzel R., Gellert M. Regulation of the genes for E. coli DNA gyrase: homeostatic control of DNA supercoiling. Cell. 1983 Aug;34(1):105–113. doi: 10.1016/0092-8674(83)90140-x. [DOI] [PubMed] [Google Scholar]
  15. Nelson H. C., Finch J. T., Luisi B. F., Klug A. The structure of an oligo(dA).oligo(dT) tract and its biological implications. Nature. 1987 Nov 19;330(6145):221–226. doi: 10.1038/330221a0. [DOI] [PubMed] [Google Scholar]
  16. Roe J. H., Burgess R. R., Record M. T., Jr Kinetics and mechanism of the interaction of Escherichia coli RNA polymerase with the lambda PR promoter. J Mol Biol. 1984 Jul 15;176(4):495–522. doi: 10.1016/0022-2836(84)90174-8. [DOI] [PubMed] [Google Scholar]
  17. Sinha N. D., Biernat J., McManus J., Köster H. Polymer support oligonucleotide synthesis XVIII: use of beta-cyanoethyl-N,N-dialkylamino-/N-morpholino phosphoramidite of deoxynucleosides for the synthesis of DNA fragments simplifying deprotection and isolation of the final product. Nucleic Acids Res. 1984 Jun 11;12(11):4539–4557. doi: 10.1093/nar/12.11.4539. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Stefano J. E., Gralla J. D. Spacer mutations in the lac ps promoter. Proc Natl Acad Sci U S A. 1982 Feb;79(4):1069–1072. doi: 10.1073/pnas.79.4.1069. [DOI] [PMC free article] [PubMed] [Google Scholar]

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