Skip to main content
PMC home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1985 May 10;13(9):3147–3159. doi: 10.1093/nar/13.9.3147

Diphtheria toxin promoter function in Corynebacterium diphtheriae and Escherichia coli.

M Kaczorek, G Zettlmeissl, F Delpeyroux, R E Streeck
PMCID: PMC341226  PMID: 3923442

Abstract

The expression of the diphtheria tox228 gene encoding the nontoxic, serologically related CRM228 mutant diphtheria toxin has been analyzed in Corynebacterium diphtheriae and Escherichia coli. The diphtheria toxin promoter has been used to direct the expression of beta-galactosidase in E.coli, and the efficiency of promotion has been compared to that obtained with the lac promoter. Expression in C.diphtheriae is known to be dependent on the absence of iron, and we present for the first time direct evidence that this regulation occurs at the level of transcription. The 5' end of toxin mRNA maps at the same position in C.diphtheriae and E.coli, suggesting identical sequences to be recognized by C.diphtheriae and E.coli RNA polymerase. The diphtheria toxin promoter carries at position -34 a TTGATT sequence closely related to the E.coli -35 consensus sequence and in the -14 to -8 region a set of overlapping sequences with complete or partial homology to the E.coli -10 consensus sequence.

Full text

PDF
3147

Images in this article

3151Image
on p.3151
3153Image
on p.3153
3154Image
on p.3154

Selected References

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

  1. Barry G., Squires C., Squires C. L. Attenuation and processing of RNA from the rplJL--rpoBC transcription unit of Escherichia coli. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3331–3335. doi: 10.1073/pnas.77.6.3331. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Charnay P., Gervais M., Louise A., Galibert F., Tiollais P. Biosynthesis of hepatitis B virus surface antigen in Escherichia coli. Nature. 1980 Aug 28;286(5776):893–895. doi: 10.1038/286893a0. [DOI] [PubMed] [Google Scholar]
  3. Giannini G., Rappuoli R., Ratti G. The amino-acid sequence of two non-toxic mutants of diphtheria toxin: CRM45 and CRM197. Nucleic Acids Res. 1984 May 25;12(10):4063–4069. doi: 10.1093/nar/12.10.4063. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Greenfield L., Bjorn M. J., Horn G., Fong D., Buck G. A., Collier R. J., Kaplan D. A. Nucleotide sequence of the structural gene for diphtheria toxin carried by corynebacteriophage beta. Proc Natl Acad Sci U S A. 1983 Nov;80(22):6853–6857. doi: 10.1073/pnas.80.22.6853. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Ishii-Kanei C., Uchida T., Yoneda M. Mutants of Corynebacterium diphtheriae PW8 that produce toxin in medium with excess iron. Appl Environ Microbiol. 1981 Dec;42(6):1130–1131. doi: 10.1128/aem.42.6.1130-1131.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Kaczorek M., Delpeyroux F., Chenciner N., Streeck R. E., Murphy J. R., Boquet P., Tiollais P. Nucleotide sequence and expression of the diphtheria tox228 gene in Escherichia coli. Science. 1983 Aug 26;221(4613):855–858. doi: 10.1126/science.6348945. [DOI] [PubMed] [Google Scholar]
  7. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  8. Leong D., Coleman K. D., Murphy J. R. Cloned fragment A of diphtheria toxin is expressed and secreted into the periplasmic space of Escherichia coli K12. Science. 1983 Apr 29;220(4596):515–517. doi: 10.1126/science.6403984. [DOI] [PubMed] [Google Scholar]
  9. Losick R., Pero J. Cascades of Sigma factors. Cell. 1981 Sep;25(3):582–584. doi: 10.1016/0092-8674(81)90164-1. [DOI] [PubMed] [Google Scholar]
  10. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  11. McLaughlin J. R., Murray C. L., Rabinowitz J. C. Unique features in the ribosome binding site sequence of the gram-positive Staphylococcus aureus beta-lactamase gene. J Biol Chem. 1981 Nov 10;256(21):11283–11291. [PubMed] [Google Scholar]
  12. Mekalanos J. J., Swartz D. J., Pearson G. D., Harford N., Groyne F., de Wilde M. Cholera toxin genes: nucleotide sequence, deletion analysis and vaccine development. Nature. 1983 Dec 8;306(5943):551–557. doi: 10.1038/306551a0. [DOI] [PubMed] [Google Scholar]
  13. Murphy J. R., Michel J. L., Teng M. Evidence that the regulation of diphtheria toxin production is directed at the level of transcription. J Bacteriol. 1978 Aug;135(2):511–516. doi: 10.1128/jb.135.2.511-516.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Murphy J. R., Pappenheimer A. M., Jr, de Borms S. T. Synthesis of diphtheria tox-gene products in Escherichia coli extracts. Proc Natl Acad Sci U S A. 1974 Jan;71(1):11–15. doi: 10.1073/pnas.71.1.11. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Murphy J. R., Skiver J., McBride G. Isolation and partial characterization of a corynebacteriophage beta, tox operator constitutive-like mutant lysogen of Corynebacterium diphtheriae. J Virol. 1976 Apr;18(1):235–244. doi: 10.1128/jvi.18.1.235-244.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Pappenheimer A. M., Jr Diphtheria toxin. Annu Rev Biochem. 1977;46:69–94. doi: 10.1146/annurev.bi.46.070177.000441. [DOI] [PubMed] [Google Scholar]
  17. Ratti G., Rappuoli R., Giannini G. The complete nucleotide sequence of the gene coding for diphtheria toxin in the corynephage omega (tox+) genome. Nucleic Acids Res. 1983 Oct 11;11(19):6589–6595. doi: 10.1093/nar/11.19.6589. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Shapira S. K., Chou J., Richaud F. V., Casadaban M. J. New versatile plasmid vectors for expression of hybrid proteins coded by a cloned gene fused to lacZ gene sequences encoding an enzymatically active carboxy-terminal portion of beta-galactosidase. Gene. 1983 Nov;25(1):71–82. doi: 10.1016/0378-1119(83)90169-5. [DOI] [PubMed] [Google Scholar]
  19. Sollner-Webb B., Reeder R. H. The nucleotide sequence of the initiation and termination sites for ribosomal RNA transcription in X. laevis. Cell. 1979 Oct;18(2):485–499. doi: 10.1016/0092-8674(79)90066-7. [DOI] [PubMed] [Google Scholar]
  20. Uchida T., Pappenheimer A. M., Jr, Harper A. A. Diphtheria toxin and related proteins. 3. Reconstitution of hybrid "diphtheria toxin" from nontoxic mutant proteins. J Biol Chem. 1973 Jun 10;248(11):3851–3854. [PubMed] [Google Scholar]
  21. Welkos S. L., Holmes R. K. Regulation of toxinogenesis in Corynebacterium diphtheriae. II. Genetic mapping of a tox regulatory mutation in bacteriophage beta. J Virol. 1981 Mar;37(3):946–954. doi: 10.1128/jvi.37.3.946-954.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. von Hippel P. H., Bear D. G., Morgan W. D., McSwiggen J. A. Protein-nucleic acid interactions in transcription: a molecular analysis. Annu Rev Biochem. 1984;53:389–446. doi: 10.1146/annurev.bi.53.070184.002133. [DOI] [PubMed] [Google Scholar]

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

RESOURCES