Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1988 Aug 11;16(15):7269–7285. doi: 10.1093/nar/16.15.7269

Transcription initiation at the tet promoter and effect of mutations.

C B Harley 1, J Lawrie 1, M Betlach 1, R Crea 1, H W Boyer 1, J Hedgpeth 1
PMCID: PMC338408  PMID: 3045754

Abstract

We have identified the startpoint for transcription in vitro of the tetracycline resistance gene (tet) of pBR322 and several deletion and insertion mutations which alter tet promoter structure. Tetracycline resistance in host bacteria correlates qualitatively with the efficiency of DNA fragments from these plasmids to promote tet transcription in vitro. Only in active promoters could we find by computer analysis promoter structures in which the -10 and -35 sequences and the relative spacing of the two regions agree with consensus sequence determinants. These data support the current model of the E. coli promoter sequence. Two promoter mutants gave heterogeneous 5' termini with additional A residues not encoded by the DNA sequence.

Full text

PDF
7269

Images in this article

7273Image
on p.7273
7274Image
on p.7274
7275Image
on p.7275
7279Image
on p.7279

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Belin D., Hedgpeth J., Selzer G. B., Epstein R. H. Temperature-sensitive mutation in the initiation codon of the rIIB gene of bacteriophage T4. Proc Natl Acad Sci U S A. 1979 Feb;76(2):700–704. doi: 10.1073/pnas.76.2.700. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bertrand-Burggraf E., Schnarr M., Lefevre J. F., Daune M. Effect of superhelicity on the transcription from the tet promoter of pBR322. Abortive initiation and unwinding experiments. Nucleic Acids Res. 1984 Oct 25;12(20):7741–7752. doi: 10.1093/nar/12.20.7741. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Blake K. R., Murakami A., Spitz S. A., Glave S. A., Reddy M. P., Ts'o P. O., Miller P. S. Hybridization arrest of globin synthesis in rabbit reticulocyte lysates and cells by oligodeoxyribonucleoside methylphosphonates. Biochemistry. 1985 Oct 22;24(22):6139–6145. doi: 10.1021/bi00343a016. [DOI] [PubMed] [Google Scholar]
  4. Brosius J., Cate R. L., Perlmutter A. P. Precise location of two promoters for the beta-lactamase gene of pBR322. S1 mapping of ribonucleic acid isolated from Escherichia coli or synthesized in vitro. J Biol Chem. 1982 Aug 10;257(15):9205–9210. [PubMed] [Google Scholar]
  5. Chan P. T., Lebowitz J., Bastia D. Nucleotide sequence determination of a strong promoter of the colicin E 1 plasmid. Analysis of restriction sites protected by RNA polymerase interactions before and after limited transcription. Nucleic Acids Res. 1979 Nov 10;7(5):1247–1262. doi: 10.1093/nar/7.5.1247. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Coleman J., Green P. J., Inouye M. The use of RNAs complementary to specific mRNAs to regulate the expression of individual bacterial genes. Cell. 1984 Jun;37(2):429–436. doi: 10.1016/0092-8674(84)90373-8. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Harley C. B., Reynolds R. P. Analysis of E. coli promoter sequences. Nucleic Acids Res. 1987 Mar 11;15(5):2343–2361. doi: 10.1093/nar/15.5.2343. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Harr R., Häggström M., Gustafsson P. Search algorithm for pattern match analysis of nucleic acid sequences. Nucleic Acids Res. 1983 May 11;11(9):2943–2957. doi: 10.1093/nar/11.9.2943. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. Hedgpeth J., Clement J. M., Marchal C., Perrin D., Hofnung M. DNA sequence encoding the NH2-terminal peptide involved in transport of lambda receptor, an Escherichia coli secretory protein. Proc Natl Acad Sci U S A. 1980 May;77(5):2621–2625. doi: 10.1073/pnas.77.5.2621. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Heyden B., Nüsslein C., Schaller H. Initiation of transcription within an RNA-polymerase binding site. Eur J Biochem. 1975 Jun 16;55(1):147–155. doi: 10.1111/j.1432-1033.1975.tb02147.x. [DOI] [PubMed] [Google Scholar]
  13. Hirose S., Suzuki Y. In vitro transcription of eukaryotic genes is affected differently by the degree of DNA supercoiling. Proc Natl Acad Sci U S A. 1988 Feb;85(3):718–722. doi: 10.1073/pnas.85.3.718. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Johnson L. D., Lazzarini R. A. The 5' terminal nucleotide of RNA from vesicular stomatitis virus defective interfering particles. Virology. 1977 Apr;77(2):863–866. doi: 10.1016/0042-6822(77)90508-6. [DOI] [PubMed] [Google Scholar]
  15. Mandecki W., Reznikoff W. S. A lac promoter with a changed distance between -10 and -35 regions. Nucleic Acids Res. 1982 Feb 11;10(3):903–912. doi: 10.1093/nar/10.3.903. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Maxam A. M., Gilbert W. A new method for sequencing DNA. Proc Natl Acad Sci U S A. 1977 Feb;74(2):560–564. doi: 10.1073/pnas.74.2.560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Miller J. H., Ippen K., Scaife J. G., Beckwith J. R. The promoter-operator region of the lac operon of Escherichia coli. J Mol Biol. 1968 Dec;38(3):413–420. doi: 10.1016/0022-2836(68)90395-1. [DOI] [PubMed] [Google Scholar]
  18. Mulligan M. E., Hawley D. K., Entriken R., McClure W. R. Escherichia coli promoter sequences predict in vitro RNA polymerase selectivity. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 2):789–800. doi: 10.1093/nar/12.1part2.789. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Peden K. W. Revised sequence of the tetracycline-resistance gene of pBR322. Gene. 1983 May-Jun;22(2-3):277–280. doi: 10.1016/0378-1119(83)90112-9. [DOI] [PubMed] [Google Scholar]
  20. Pribnow D. Nucleotide sequence of an RNA polymerase binding site at an early T7 promoter. Proc Natl Acad Sci U S A. 1975 Mar;72(3):784–788. doi: 10.1073/pnas.72.3.784. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Rodriguez R. L., West R. W., Heyneker H. L., Bolivar F., Boyer H. W. Characterizing wild-type and mutant promoters of the tetracycline resistance gene in pBR313. Nucleic Acids Res. 1979 Jul 25;6(10):3267–3287. doi: 10.1093/nar/6.10.3267. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Rosenberg M., Court D. Regulatory sequences involved in the promotion and termination of RNA transcription. Annu Rev Genet. 1979;13:319–353. doi: 10.1146/annurev.ge.13.120179.001535. [DOI] [PubMed] [Google Scholar]
  23. Sanger F., Coulson A. R. The use of thin acrylamide gels for DNA sequencing. FEBS Lett. 1978 Mar 1;87(1):107–110. doi: 10.1016/0014-5793(78)80145-8. [DOI] [PubMed] [Google Scholar]
  24. Siebenlist U., Simpson R. B., Gilbert W. E. coli RNA polymerase interacts homologously with two different promoters. Cell. 1980 Jun;20(2):269–281. doi: 10.1016/0092-8674(80)90613-3. [DOI] [PubMed] [Google Scholar]
  25. Simoncsits A., Brownlee G. G., Brown R. S., Rubin J. R., Guilley H. New rapid gel sequencing method for RNA. Nature. 1977 Oct 27;269(5631):833–836. doi: 10.1038/269833a0. [DOI] [PubMed] [Google Scholar]
  26. Staden R. Computer methods to locate signals in nucleic acid sequences. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 2):505–519. doi: 10.1093/nar/12.1part2.505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Stefano J. E., Gralla J. D. Kinetic investigation of the mechanism of RNA polymerase binding to mutant lac promoters. J Biol Chem. 1980 Nov 10;255(21):10423–10430. [PubMed] [Google Scholar]
  28. 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]
  29. Stüber D., Bujard H. Organization of transcriptional signals in plasmids pBR322 and pACYC184. Proc Natl Acad Sci U S A. 1981 Jan;78(1):167–171. doi: 10.1073/pnas.78.1.167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Tait R. C., Boyer H. W. On the nature of tetracycline resistance controlled by the plasmid pSC101. Cell. 1978 Jan;13(1):73–81. doi: 10.1016/0092-8674(78)90139-3. [DOI] [PubMed] [Google Scholar]
  31. Youderian P., Bouvier S., Susskind M. M. Sequence determinants of promoter activity. Cell. 1982 Oct;30(3):843–853. doi: 10.1016/0092-8674(82)90289-6. [DOI] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES