Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1995 Apr;177(7):1896–1899. doi: 10.1128/jb.177.7.1896-1899.1995

Transcription termination in the Escherichia coli dnaA gene is not mediated by the internal DnaA box.

I Pérez-Roger 1, F Macián 1, M E Armengod 1
PMCID: PMC176824  PMID: 7896719

Abstract

DnaA protein is a DNA-binding protein which recognizes a 9-bp consensus sequence called the DnaA box. By binding to DnaA boxes, DnaA protein regulates initiation of chromosomal replication and transcription of several genes. The dnaA gene contains two DnaA boxes, one located in the regulatory region and one within the structural gene. In this paper, we explore the role of the internal DnaA box in dnaA expression because it has been proposed that the DnaA box-DnaA protein complex can block transcribing RNA polymerase. Firstly, we analyzed the degree of derepression of the dnaA gene, measured as beta-galactosidase activity of a dnaA-lacZ fusion inserted onto the bacterial chromosome, produced by an extra copy number of the dnaA DnaA boxes carried by multicopy plasmids. Secondly, we analyzed repression produced by elevated levels of DnaA protein on single-copy dnaA-lacZ fusions containing or not containing the internal DnaA box. Our results indicate that the internal DnaA box does not play a regulatory role in dnaA expression.

Full Text

The Full Text of this article is available as a PDF (188.1 KB).

Selected References

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

  1. Aldea M., Garrido T., Pla J., Vicente M. Division genes in Escherichia coli are expressed coordinately to cell septum requirements by gearbox promoters. EMBO J. 1990 Nov;9(11):3787–3794. doi: 10.1002/j.1460-2075.1990.tb07592.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Alifano P., Ciampi M. S., Nappo A. G., Bruni C. B., Carlomagno M. S. In vivo analysis of the mechanisms responsible for strong transcriptional polarity in a "sense" mutant within an intercistronic region. Cell. 1988 Oct 21;55(2):351–360. doi: 10.1016/0092-8674(88)90058-x. [DOI] [PubMed] [Google Scholar]
  3. Andrup L., Atlung T., Ogasawara N., Yoshikawa H., Hansen F. G. Interaction of the Bacillus subtilis DnaA-like protein with the Escherichia coli DnaA protein. J Bacteriol. 1988 Mar;170(3):1333–1338. doi: 10.1128/jb.170.3.1333-1338.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Armengod M. E., García-Sogo M., Lambíes E. Transcriptional organization of the dnaN and recF genes of Escherichia coli K-12. J Biol Chem. 1988 Aug 25;263(24):12109–12114. [PubMed] [Google Scholar]
  5. Armengod M. E., García-Sogo M., Pérez-Roger I., Macián F., Navarro-Aviñ J. P. Tandem transcription termination sites in the dnaN gene of Escherichia coli. J Biol Chem. 1991 Oct 15;266(29):19725–19730. [PubMed] [Google Scholar]
  6. Armengod M. E., Lambíes E. Overlapping arrangement of the recF and dnaN operons of Escherichia coli; positive and negative control sequences. Gene. 1986;43(3):183–196. doi: 10.1016/0378-1119(86)90206-4. [DOI] [PubMed] [Google Scholar]
  7. Atlung T., Clausen E. S., Hansen F. G. Autoregulation of the dnaA gene of Escherichia coli K12. Mol Gen Genet. 1985;200(3):442–450. doi: 10.1007/BF00425729. [DOI] [PubMed] [Google Scholar]
  8. Balbás P., Soberón X., Merino E., Zurita M., Lomeli H., Valle F., Flores N., Bolivar F. Plasmid vector pBR322 and its special-purpose derivatives--a review. Gene. 1986;50(1-3):3–40. doi: 10.1016/0378-1119(86)90307-0. [DOI] [PubMed] [Google Scholar]
  9. Braun R. E., O'Day K., Wright A. Autoregulation of the DNA replication gene dnaA in E. coli K-12. Cell. 1985 Jan;40(1):159–169. doi: 10.1016/0092-8674(85)90319-8. [DOI] [PubMed] [Google Scholar]
  10. Dunderdale H. J., West S. C. Recombination genes and proteins. Curr Opin Genet Dev. 1994 Apr;4(2):221–228. doi: 10.1016/s0959-437x(05)80048-6. [DOI] [PubMed] [Google Scholar]
  11. Fuller R. S., Funnell B. E., Kornberg A. The dnaA protein complex with the E. coli chromosomal replication origin (oriC) and other DNA sites. Cell. 1984 Oct;38(3):889–900. doi: 10.1016/0092-8674(84)90284-8. [DOI] [PubMed] [Google Scholar]
  12. Hansen F. G., Hansen E. B., Atlung T. The nucleotide sequence of the dnaA gene promoter and of the adjacent rpmH gene, coding for the ribosomal protein L34, of Escherichia coli. EMBO J. 1982;1(9):1043–1048. doi: 10.1002/j.1460-2075.1982.tb01294.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hansen F. G., Koefoed S., Sørensen L., Atlung T. Titration of DnaA protein by oriC DnaA-boxes increases dnaA gene expression in Escherichia coli. EMBO J. 1987 Jan;6(1):255–258. doi: 10.1002/j.1460-2075.1987.tb04747.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kücherer C., Lother H., Kölling R., Schauzu M. A., Messer W. Regulation of transcription of the chromosomal dnaA gene of Escherichia coli. Mol Gen Genet. 1986 Oct;205(1):115–121. doi: 10.1007/BF02428040. [DOI] [PubMed] [Google Scholar]
  15. Lloyd R. G., Sharples G. J. Genetic analysis of recombination in prokaryotes. Curr Opin Genet Dev. 1992 Oct;2(5):683–690. doi: 10.1016/s0959-437x(05)80127-3. [DOI] [PubMed] [Google Scholar]
  16. Lother H., Kölling R., Kücherer C., Schauzu M. dnaA protein-regulated transcription: effects on the in vitro replication of Escherichia coli minichromosomes. EMBO J. 1985 Feb;4(2):555–560. doi: 10.1002/j.1460-2075.1985.tb03664.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Løbner-Olesen A., Skarstad K., Hansen F. G., von Meyenburg K., Boye E. The DnaA protein determines the initiation mass of Escherichia coli K-12. Cell. 1989 Jun 2;57(5):881–889. doi: 10.1016/0092-8674(89)90802-7. [DOI] [PubMed] [Google Scholar]
  18. Macián F., Pérez-Roger I., Armengod M. E. An improved vector system for constructing transcriptional lacZ fusions: analysis of regulation of the dnaA, dnaN, recF and gyrB genes of Escherichia coli. Gene. 1994 Jul 22;145(1):17–24. doi: 10.1016/0378-1119(94)90317-4. [DOI] [PubMed] [Google Scholar]
  19. Maki S., Kornberg A. DNA polymerase III holoenzyme of Escherichia coli. III. Distinctive processive polymerases reconstituted from purified subunits. J Biol Chem. 1988 May 15;263(14):6561–6569. [PubMed] [Google Scholar]
  20. Masters M., Paterson T., Popplewell A. G., Owen-Hughes T., Pringle J. H., Begg K. J. The effect of DnaA protein levels and the rate of initiation at oriC on transcription originating in the ftsQ and ftsA genes: in vivo experiments. Mol Gen Genet. 1989 Apr;216(2-3):475–483. doi: 10.1007/BF00334393. [DOI] [PubMed] [Google Scholar]
  21. Matsumoto Y., Shigesada K., Hirano M., Imai M. Autogenous regulation of the gene for transcription termination factor rho in Escherichia coli: localization and function of its attenuators. J Bacteriol. 1986 Jun;166(3):945–958. doi: 10.1128/jb.166.3.945-958.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Ostrow K. S., Silhavy T. J., Garrett S. cis-acting sites required for osmoregulation of ompF expression in Escherichia coli K-12. J Bacteriol. 1986 Dec;168(3):1165–1171. doi: 10.1128/jb.168.3.1165-1171.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Pérez-Roger I., García-Sogo M., Navarro-Aviñ J. P., López-Acedo C., Macián F., Armengod M. E. Positive and negative regulatory elements in the dnaA-dnaN-recF operon of Escherichia coli. Biochimie. 1991 Feb-Mar;73(2-3):329–334. doi: 10.1016/0300-9084(91)90220-u. [DOI] [PubMed] [Google Scholar]
  24. Ruteshouser E. C., Richardson J. P. Identification and characterization of transcription termination sites in the Escherichia coli lacZ gene. J Mol Biol. 1989 Jul 5;208(1):23–43. doi: 10.1016/0022-2836(89)90085-5. [DOI] [PubMed] [Google Scholar]
  25. Schaefer C., Messer W. Directionality of DnaA protein/DNA interaction. Active orientation of the DnaA protein/dnaA box complex in transcription termination. EMBO J. 1989 May;8(5):1609–1613. doi: 10.1002/j.1460-2075.1989.tb03545.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Schaefer C., Messer W. Termination of the Escherichia coli asnC transcript. The DnaA protein/dnaA box complex blocks transcribing RNA polymerase. Gene. 1988 Dec 20;73(2):347–354. doi: 10.1016/0378-1119(88)90499-4. [DOI] [PubMed] [Google Scholar]
  27. Slauch J. M., Silhavy T. J. Genetic fusions as experimental tools. Methods Enzymol. 1991;204:213–248. doi: 10.1016/0076-6879(91)04011-c. [DOI] [PubMed] [Google Scholar]
  28. Solomon K. A., Hsu D. K., Brusilow W. S. Use of lacZ fusions to measure in vivo expression of the first three genes of the Escherichia coli unc operon. J Bacteriol. 1989 Jun;171(6):3039–3045. doi: 10.1128/jb.171.6.3039-3045.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Stanssens P., Remaut E., Fiers W. Inefficient translation initiation causes premature transcription termination in the lacZ gene. Cell. 1986 Mar 14;44(5):711–718. doi: 10.1016/0092-8674(86)90837-8. [DOI] [PubMed] [Google Scholar]
  30. Tesfa-Selase F., Drabble W. T. Regulation of the gua operon of Escherichia coli by the DnaA protein. Mol Gen Genet. 1992 Jan;231(2):256–264. doi: 10.1007/BF00279799. [DOI] [PubMed] [Google Scholar]
  31. Wang Q. P., Kaguni J. M. Transcriptional repression of the dnaA gene of Escherichia coli by dnaA protein. Mol Gen Genet. 1987 Oct;209(3):518–525. doi: 10.1007/BF00331158. [DOI] [PubMed] [Google Scholar]
  32. Wang Q. P., Kaguni J. M. dnaA protein regulates transcriptions of the rpoH gene of Escherichia coli. J Biol Chem. 1989 May 5;264(13):7338–7344. [PubMed] [Google Scholar]
  33. Wende M., Quinones A., Diederich L., Jueterbock W. R., Messer W. Transcription termination in the dnaA gene. Mol Gen Genet. 1991 Dec;230(3):486–490. doi: 10.1007/BF00280306. [DOI] [PubMed] [Google Scholar]
  34. Wright J. J., Hayward R. S. Transcriptional termination at a fully rho-independent site in Escherichia coli is prevented by uninterrupted translation of the nascent RNA. EMBO J. 1987 Apr;6(4):1115–1119. doi: 10.1002/j.1460-2075.1987.tb04866.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Yarchuk O., Jacques N., Guillerez J., Dreyfus M. Interdependence of translation, transcription and mRNA degradation in the lacZ gene. J Mol Biol. 1992 Aug 5;226(3):581–596. doi: 10.1016/0022-2836(92)90617-s. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES