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. 1990 Nov 11;18(21):6271–6275. doi: 10.1093/nar/18.21.6271

The signal for growth rate control and stringent sensitivity in E. coli is not restricted to a particular sequence motif within the promoter region.

M Zacharias 1, H U Göringer 1, R Wagner 1
PMCID: PMC332491  PMID: 2243774

Abstract

Hybrid promoter constructs were used to determine the DNA sequence requirements for stringent and growth rate control within a promoter region. The promoters were obtained by fusing complementing sequence regions located upstream and downstream from the GCGC discriminator motif of the growth rate regulated rRNA P1 promoter and a non-regulated tac promoter variant. The activities and the regulatory response of the hybrid promoters were determined in vivo using a promoter test vector system with the chloramphenicol acetyltransferase (CAT) reporter gene. Measurements were made at different growth rates and after starvation for isoleucine to induce the stringent response. Neither the upstream nor the downstream sequence of P1 relative to the GCGC discriminator motif conferred comparable regulatory features when fused to the complementing sequences of the non-regulated mutant tac promoter. A minor response to amino acid deprivation or changes in the growth rate was noted for the hybrid promoter with the rrnB P1 upstream segment and the tac downstream element, pointing to a slightly different importance of the two sequence elements for regulation. The parallel effects for stringent as well as growth rate regulation of the hybrid promoters supports the view of a common mechanism for both types of control. However, none of the promoter sequence elements on its own was able to restore the complete regulatory behaviour of their 'parent' promoters.

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Selected References

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

  1. Baracchini E., Bremer H. Stringent and growth control of rRNA synthesis in Escherichia coli are both mediated by ppGpp. J Biol Chem. 1988 Feb 25;263(6):2597–2602. [PubMed] [Google Scholar]
  2. Brosius J. Plasmid vectors for the selection of promoters. Gene. 1984 Feb;27(2):151–160. doi: 10.1016/0378-1119(84)90136-7. [DOI] [PubMed] [Google Scholar]
  3. Deneer H. G., Spiegelman G. B. Bacillus subtilis rRNA promoters are growth rate regulated in Escherichia coli. J Bacteriol. 1987 Mar;169(3):995–1002. doi: 10.1128/jb.169.3.995-1002.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dickson R. R., Gaal T., deBoer H. A., deHaseth P. L., Gourse R. L. Identification of promoter mutants defective in growth-rate-dependent regulation of rRNA transcription in Escherichia coli. J Bacteriol. 1989 Sep;171(9):4862–4870. doi: 10.1128/jb.171.9.4862-4870.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Duester G., Elford R. M., Holmes W. M. Fusion of the Escherichia coli tRNALeu1 promoter to the galK gene: analysis of sequences necessary for growth-rate-dependent regulation. Cell. 1982 Oct;30(3):855–864. doi: 10.1016/0092-8674(82)90290-2. [DOI] [PubMed] [Google Scholar]
  6. Feinberg A. P., Vogelstein B. "A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity". Addendum. Anal Biochem. 1984 Feb;137(1):266–267. doi: 10.1016/0003-2697(84)90381-6. [DOI] [PubMed] [Google Scholar]
  7. Fiil N., Friesen J. D. Isolation of "relaxed" mutants of Escherichia coli. J Bacteriol. 1968 Feb;95(2):729–731. doi: 10.1128/jb.95.2.729-731.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gausing K. Regulation of ribosome production in Escherichia coli: synthesis and stability of ribosomal RNA and of ribosomal protein messenger RNA at different growth rates. J Mol Biol. 1977 Sep 25;115(3):335–354. doi: 10.1016/0022-2836(77)90158-9. [DOI] [PubMed] [Google Scholar]
  9. Gorman C. M., Moffat L. F., Howard B. H. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol. 1982 Sep;2(9):1044–1051. doi: 10.1128/mcb.2.9.1044. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gourse R. L., de Boer H. A., Nomura M. DNA determinants of rRNA synthesis in E. coli: growth rate dependent regulation, feedback inhibition, upstream activation, antitermination. Cell. 1986 Jan 17;44(1):197–205. doi: 10.1016/0092-8674(86)90498-8. [DOI] [PubMed] [Google Scholar]
  11. Igarashi K., Fujita N., Ishihama A. Promoter selectivity of Escherichia coli RNA polymerase: omega factor is responsible for the ppGpp sensitivity. Nucleic Acids Res. 1989 Nov 11;17(21):8755–8765. doi: 10.1093/nar/17.21.8755. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Klotsky R. A., Schwartz I. Measurement of cat expression from growth-rate-regulated promoters employing beta-lactamase activity as an indicator of plasmid copy number. Gene. 1987;55(1):141–146. doi: 10.1016/0378-1119(87)90257-5. [DOI] [PubMed] [Google Scholar]
  13. Lamond A. I., Travers A. A. Genetically separable functional elements mediate the optimal expression and stringent regulation of a bacterial tRNA gene. Cell. 1985 Feb;40(2):319–326. doi: 10.1016/0092-8674(85)90146-1. [DOI] [PubMed] [Google Scholar]
  14. Lamond A. I., Travers A. A. Stringent control of bacterial transcription. Cell. 1985 May;41(1):6–8. doi: 10.1016/0092-8674(85)90050-9. [DOI] [PubMed] [Google Scholar]
  15. Little R., Ryals J., Bremer H. rpoB mutation in Escherichia coli alters control of ribosome synthesis by guanosine tetraphosphate. J Bacteriol. 1983 May;154(2):787–792. doi: 10.1128/jb.154.2.787-792.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lupski J. R., Ruiz A. A., Godson G. N. Promotion, termination, and anti-termination in the rpsU-dnaG-rpoD macromolecular synthesis operon of E. coli K-12. Mol Gen Genet. 1984;195(3):391–401. doi: 10.1007/BF00341439. [DOI] [PubMed] [Google Scholar]
  17. Mizushima-Sugano J., Kaziro Y. Regulation of the expression of the tufB operon: DNA sequences directly involved in the stringent control. EMBO J. 1985 Apr;4(4):1053–1058. doi: 10.1002/j.1460-2075.1985.tb03738.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Ryals J., Bremer H. relA-dependent RNA polymerase activity in Escherichia coli. J Bacteriol. 1982 Apr;150(1):168–179. doi: 10.1128/jb.150.1.168-179.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Sarubbi E., Rudd K. E., Cashel M. Basal ppGpp level adjustment shown by new spoT mutants affect steady state growth rates and rrnA ribosomal promoter regulation in Escherichia coli. Mol Gen Genet. 1988 Aug;213(2-3):214–222. doi: 10.1007/BF00339584. [DOI] [PubMed] [Google Scholar]
  21. Travers A. A., Lamond A. I., Weeks J. R. Alteration of the growth-rate-dependent regulation of Escherichia coli tyrT expression by promoter mutations. J Mol Biol. 1986 May 5;189(1):251–255. doi: 10.1016/0022-2836(86)90397-9. [DOI] [PubMed] [Google Scholar]
  22. Travers A. A. Promoter sequence for stringent control of bacterial ribonucleic acid synthesis. J Bacteriol. 1980 Feb;141(2):973–976. doi: 10.1128/jb.141.2.973-976.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Travers A. Modulation of RNA polymerase specificity by ppGpp. Mol Gen Genet. 1976 Aug 19;147(2):225–232. doi: 10.1007/BF00267575. [DOI] [PubMed] [Google Scholar]
  24. Zacharias M., Göringer H. U., Wagner R. Influence of the GCGC discriminator motif introduced into the ribosomal RNA P2- and tac promoter on growth-rate control and stringent sensitivity. EMBO J. 1989 Nov;8(11):3357–3363. doi: 10.1002/j.1460-2075.1989.tb08498.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Zacharias M., Wagner R. Functional characterization of a putative internal promoter sequence between the 16S and the 23S RNA genes within the Escherichia coli rrnB operon. Mol Microbiol. 1989 Mar;3(3):405–410. doi: 10.1111/j.1365-2958.1989.tb00185.x. [DOI] [PubMed] [Google Scholar]

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