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. 1989 Nov;171(11):5955–5962. doi: 10.1128/jb.171.11.5955-5962.1989

Regulation of expression of the Pasteurella haemolytica leukotoxin determinant.

C A Strathdee 1, R Y Lo 1
PMCID: PMC210460  PMID: 2478522

Abstract

The Pasteurella haemolytica leukotoxin determinant is composed of four contiguous genes encoded on the same DNA strand and denoted lktCABD, in the order of their genetic organization. To gain a better understanding of the expression and regulation of the leukotoxin, the transcripts and promoters of the lkt determinant were mapped. Northern (RNA) blot analysis revealed two sets of transcripts. One set was 3.7 and 3.4 kilobases long, encoded lktCA, and comprised approximately 90% of the transcripts, whereas the other set was 7.4 and 7.1 kilobases long and encoded lktCABD. Two promoters were present, and each had features similar to the Escherichia coli consensus promoter sequences. Both promoters were located upstream from lktC; they were separated by 258 base pairs, as mapped by primer extension analysis. These results suggest a mechanism of expression similar to that of the related E. coli hemolysin. Transcription initiated upstream from lktC at either promoter and continued through lktC and lktA to a rho-independent transcriptional termination signal in the lktA-lktB intercistronic region. This signal attenuated expression by terminating 90% of transcription to generate the 3.7- and 3.4-kilobase lktCA transcripts. The remaining readthrough transcription generated full-length 7.4- and 7.1-kilobase lktCABD transcripts. Expression of the leukotoxin was greatly reduced by growth at 30 degrees C, pH 6.5, and Fe2+ limitation. These conditions also modulated the expression of a number of other secreted proteins, which suggests that all of these secreted proteins are controlled by the same regulatory mechanism.

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Selected References

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

  1. Aiba H., Adhya S., de Crombrugghe B. Evidence for two functional gal promoters in intact Escherichia coli cells. J Biol Chem. 1981 Nov 25;256(22):11905–11910. [PubMed] [Google Scholar]
  2. Bjorn M. J., Sokol P. A., Iglewski B. H. Influence of iron on yields of extracellular products in Pseudomonas aeruginosa cultures. J Bacteriol. 1979 Apr;138(1):193–200. doi: 10.1128/jb.138.1.193-200.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Calderwood S. B., Mekalanos J. J. Iron regulation of Shiga-like toxin expression in Escherichia coli is mediated by the fur locus. J Bacteriol. 1987 Oct;169(10):4759–4764. doi: 10.1128/jb.169.10.4759-4764.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Felmlee T., Pellett S., Welch R. A. Nucleotide sequence of an Escherichia coli chromosomal hemolysin. J Bacteriol. 1985 Jul;163(1):94–105. doi: 10.1128/jb.163.1.94-105.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Frank D. W., Iglewski B. H. Kinetics of toxA and regA mRNA accumulation in Pseudomonas aeruginosa. J Bacteriol. 1988 Oct;170(10):4477–4483. doi: 10.1128/jb.170.10.4477-4483.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Frank G. H., Tabatabai L. B. Neuraminidase activity of Pasteurella haemolytica isolates. Infect Immun. 1981 Jun;32(3):1119–1122. doi: 10.1128/iai.32.3.1119-1122.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fujita N., Ishihama A. Heat-shock induction of RNA polymerase sigma-32 synthesis in Escherichia coli: transcriptional control and a multiple promoter system. Mol Gen Genet. 1987 Nov;210(1):10–15. doi: 10.1007/BF00337752. [DOI] [PubMed] [Google Scholar]
  8. Gentry M. J., Confer A. W., Weinberg E. D., Homer J. T. Cytotoxin (leukotoxin) production by Pasteurella haemolytica: requirement for an iron-containing compound. Am J Vet Res. 1986 Sep;47(9):1919–1923. [PubMed] [Google Scholar]
  9. Glaser P., Ladant D., Sezer O., Pichot F., Ullmann A., Danchin A. The calmodulin-sensitive adenylate cyclase of Bordetella pertussis: cloning and expression in Escherichia coli. Mol Microbiol. 1988 Jan;2(1):19–30. [PubMed] [Google Scholar]
  10. Griggs D. W., Tharp B. B., Konisky J. Cloning and promoter identification of the iron-regulated cir gene of Escherichia coli. J Bacteriol. 1987 Dec;169(12):5343–5352. doi: 10.1128/jb.169.12.5343-5352.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hagblom P., Segal E., Billyard E., So M. Intragenic recombination leads to pilus antigenic variation in Neisseria gonorrhoeae. Nature. 1985 May 9;315(6015):156–158. doi: 10.1038/315156a0. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Juarez A., Hughes C., Vogel M., Goebel W. Expression and regulation of the plasmid-encoded hemolysin determinant of Escherichia coli. Mol Gen Genet. 1984;197(2):196–203. doi: 10.1007/BF00330963. [DOI] [PubMed] [Google Scholar]
  14. Kajitani M., Ishihama A. Promoter selectivity of Escherichia coli RNA polymerase. Differential stringent control of the multiple promoters from ribosomal RNA and protein operons. J Biol Chem. 1984 Feb 10;259(3):1951–1957. [PubMed] [Google Scholar]
  15. Knapp S., Mekalanos J. J. Two trans-acting regulatory genes (vir and mod) control antigenic modulation in Bordetella pertussis. J Bacteriol. 1988 Nov;170(11):5059–5066. doi: 10.1128/jb.170.11.5059-5066.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kolter R., Yanofsky C. Attenuation in amino acid biosynthetic operons. Annu Rev Genet. 1982;16:113–134. doi: 10.1146/annurev.ge.16.120182.000553. [DOI] [PubMed] [Google Scholar]
  17. Koronakis V., Cross M., Hughes C. Expression of the E.coli hemolysin secretion gene hlyB involves transcript anti-termination within the hly operon. Nucleic Acids Res. 1988 Jun 10;16(11):4789–4800. doi: 10.1093/nar/16.11.4789. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Koronakis V., Cross M., Senior B., Koronakis E., Hughes C. The secreted hemolysins of Proteus mirabilis, Proteus vulgaris, and Morganella morganii are genetically related to each other and to the alpha-hemolysin of Escherichia coli. J Bacteriol. 1987 Apr;169(4):1509–1515. doi: 10.1128/jb.169.4.1509-1515.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Koronakis V., Hughes C. Identification of the promoters directing in vivo expression of hemolysin genes in Proteus vulgaris and Escherichia coli. Mol Gen Genet. 1988 Jul;213(1):99–104. doi: 10.1007/BF00333404. [DOI] [PubMed] [Google Scholar]
  20. Lo R. Y., Strathdee C. A., Shewen P. E. Nucleotide sequence of the leukotoxin genes of Pasteurella haemolytica A1. Infect Immun. 1987 Sep;55(9):1987–1996. doi: 10.1128/iai.55.9.1987-1996.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Mackman N., Nicaud J. M., Gray L., Holland I. B. Identification of polypeptides required for the export of haemolysin 2001 from E. coli. Mol Gen Genet. 1985;201(3):529–536. doi: 10.1007/BF00331351. [DOI] [PubMed] [Google Scholar]
  22. Martin S. W., Meek A. H., Davis D. G., Thomson R. G., Johnson J. A., Lopez A., Stephens L., Curtis R. A., Prescott J. F., Rosendal S. Factors associated with mortality in feedlot cattle: the Bruce County Beef Cattle Project. Can J Comp Med. 1980 Jan;44(1):1–10. [PMC free article] [PubMed] [Google Scholar]
  23. McMaster G. K., Carmichael G. G. Analysis of single- and double-stranded nucleic acids on polyacrylamide and agarose gels by using glyoxal and acridine orange. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4835–4838. doi: 10.1073/pnas.74.11.4835. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Nicaud J. M., Mackman N., Gray L., Holland I. B. Characterisation of HlyC and mechanism of activation and secretion of haemolysin from E. coli 2001. FEBS Lett. 1985 Aug 5;187(2):339–344. doi: 10.1016/0014-5793(85)81272-2. [DOI] [PubMed] [Google Scholar]
  25. Otulakowski G. L., Shewen P. E., Udoh A. E., Mellors A., Wilkie B. N. Proteolysis of sialoglycoprotein by Pasteurella haemolytica cytotoxic culture supernatant. Infect Immun. 1983 Oct;42(1):64–70. doi: 10.1128/iai.42.1.64-70.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Peterson K. M., Mekalanos J. J. Characterization of the Vibrio cholerae ToxR regulon: identification of novel genes involved in intestinal colonization. Infect Immun. 1988 Nov;56(11):2822–2829. doi: 10.1128/iai.56.11.2822-2829.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  28. Rogers H. J. Iron-Binding Catechols and Virulence in Escherichia coli. Infect Immun. 1973 Mar;7(3):445–456. doi: 10.1128/iai.7.3.445-456.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Strathdee C. A., Lo R. Y. Cloning, nucleotide sequence, and characterization of genes encoding the secretion function of the Pasteurella haemolytica leukotoxin determinant. J Bacteriol. 1989 Feb;171(2):916–928. doi: 10.1128/jb.171.2.916-928.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Tai S. P., Holmes R. K. Iron regulation of the cloned diphtheria toxin promoter in Escherichia coli. Infect Immun. 1988 Sep;56(9):2430–2436. doi: 10.1128/iai.56.9.2430-2436.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Vogel M., Hess J., Then I., Juarez A., Goebel W. Characterization of a sequence (hlyR) which enhances synthesis and secretion of hemolysin in Escherichia coli. Mol Gen Genet. 1988 Apr;212(1):76–84. doi: 10.1007/BF00322447. [DOI] [PubMed] [Google Scholar]
  32. Wagner W., Vogel M., Goebel W. Transport of hemolysin across the outer membrane of Escherichia coli requires two functions. J Bacteriol. 1983 Apr;154(1):200–210. doi: 10.1128/jb.154.1.200-210.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Welch R. A. Identification of two different hemolysin determinants in uropathogenic Proteus isolates. Infect Immun. 1987 Sep;55(9):2183–2190. doi: 10.1128/iai.55.9.2183-2190.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Welch R. A., Pellett S. Transcriptional organization of the Escherichia coli hemolysin genes. J Bacteriol. 1988 Apr;170(4):1622–1630. doi: 10.1128/jb.170.4.1622-1630.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Yanofsky C. Attenuation in the control of expression of bacterial operons. Nature. 1981 Feb 26;289(5800):751–758. doi: 10.1038/289751a0. [DOI] [PubMed] [Google Scholar]
  36. Yates W. D. A review of infectious bovine rhinotracheitis, shipping fever pneumonia and viral-bacterial synergism in respiratory disease of cattle. Can J Comp Med. 1982 Jul;46(3):225–263. [PMC free article] [PubMed] [Google Scholar]
  37. de Lorenzo V., Wee S., Herrero M., Neilands J. B. Operator sequences of the aerobactin operon of plasmid ColV-K30 binding the ferric uptake regulation (fur) repressor. J Bacteriol. 1987 Jun;169(6):2624–2630. doi: 10.1128/jb.169.6.2624-2630.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]

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