Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1984 Jul;81(13):4120–4124. doi: 10.1073/pnas.81.13.4120

Regulation of the araC gene of Escherichia coli: catabolite repression, autoregulation, and effect on araBAD expression.

C G Miyada, L Stoltzfus, G Wilcox
PMCID: PMC345380  PMID: 6377308

Abstract

The araC gene encodes a positive regulatory protein required for L-arabinose utilization in Escherichia coli. Transcription from the araC promoter has been shown to be under positive control by cAMP receptor protein and under negative control by its protein product (autoregulation). This work describes the identification of the region of the araC promoter that interacts with the cAMP receptor protein to mediate catabolite repression. A 3-base-pair deletion centered 60 base pairs from the transcriptional initiation site results in a mutant araC promoter that, in the absence of araC protein, reduces transcriptional activity when compared with the wild-type promoter and is unresponsive to various concentrations of intracellular cAMP in vivo. The same deletion results in a lowered affinity of the araC promoter for cAMP receptor protein in vitro. However, this lowered affinity for the mutant araC promoter does not result in substantial reduction of intracellular araC protein because autoregulation of the araC gene dominates catabolite repression. The 3-base-pair deletion in the cAMP receptor protein binding site of the araC promoter does not affect catabolite repression of the adjacent araBAD operon. The implications of these results on current models for expression of the araBAD operon and the araC gene are discussed.

Full text

PDF
4120

Images in this article

4123Image
on p.4123
4123Image
on p.4123

Selected References

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

  1. Aiba H., Fujimoto S., Ozaki N. Molecular cloning and nucleotide sequencing of the gene for E. coli cAMP receptor protein. Nucleic Acids Res. 1982 Feb 25;10(4):1345–1361. doi: 10.1093/nar/10.4.1345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bachmann B. J., Low K. B. Linkage map of Escherichia coli K-12, edition 6. Microbiol Rev. 1980 Mar;44(1):1–56. doi: 10.1128/mr.44.1.1-56.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Boone T., Wilcox G. A rapid high-yield purification procedure for the cyclic adenosine 3',5'-monophosphate receptor protein from Escherichia coli. Biochim Biophys Acta. 1978 Jul 17;541(4):528–534. doi: 10.1016/0304-4165(78)90162-9. [DOI] [PubMed] [Google Scholar]
  4. Cabello F., Timmis K., Cohen S. N. Replication control in a composite plasmid constructed by in vitro linkage of two distinct replicons. Nature. 1976 Jan 29;259(5541):285–290. doi: 10.1038/259285a0. [DOI] [PubMed] [Google Scholar]
  5. Casadaban M. J., Cohen S. N. Analysis of gene control signals by DNA fusion and cloning in Escherichia coli. J Mol Biol. 1980 Apr;138(2):179–207. doi: 10.1016/0022-2836(80)90283-1. [DOI] [PubMed] [Google Scholar]
  6. Casadaban M. J. Regulation of the regulatory gene for the arabinose pathway, araC. J Mol Biol. 1976 Jul 5;104(3):557–566. doi: 10.1016/0022-2836(76)90120-0. [DOI] [PubMed] [Google Scholar]
  7. Cohen S. N., Chang A. C., Hsu L. Nonchromosomal antibiotic resistance in bacteria: genetic transformation of Escherichia coli by R-factor DNA. Proc Natl Acad Sci U S A. 1972 Aug;69(8):2110–2114. doi: 10.1073/pnas.69.8.2110. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Conner B. J., Reyes A. A., Morin C., Itakura K., Teplitz R. L., Wallace R. B. Detection of sickle cell beta S-globin allele by hybridization with synthetic oligonucleotides. Proc Natl Acad Sci U S A. 1983 Jan;80(1):278–282. doi: 10.1073/pnas.80.1.278. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Englesberg E., Wilcox G. Regulation: positive control. Annu Rev Genet. 1974;8:219–242. doi: 10.1146/annurev.ge.08.120174.001251. [DOI] [PubMed] [Google Scholar]
  10. Galas D. J., Schmitz A. DNAse footprinting: a simple method for the detection of protein-DNA binding specificity. Nucleic Acids Res. 1978 Sep;5(9):3157–3170. doi: 10.1093/nar/5.9.3157. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gielow L., Largen M., Englesberg E. Initiator constitutive mutants of the L-arabinose operon (OIBAD) of Escherichia coli B/r. Genetics. 1971 Nov;69(3):289–302. doi: 10.1093/genetics/69.3.289. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Greenfield L., Boone T., Wilcox G. DNA sequence of the araBAD promoter in Escherichia coli B/r. Proc Natl Acad Sci U S A. 1978 Oct;75(10):4724–4728. doi: 10.1073/pnas.75.10.4724. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hahn S., Schleif R. In vivo regulation of the Escherichia coli araC promoter. J Bacteriol. 1983 Aug;155(2):593–600. doi: 10.1128/jb.155.2.593-600.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Heffernan L., Bass R., Englesberg E. Mutations affecting catabolite repression of the L-arabinose regulon in Escherichia coli B/r. J Bacteriol. 1976 Jun;126(3):1119–1131. doi: 10.1128/jb.126.3.1119-1131.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hirsh J., Schleif R. The araC promoter: transcription, mapping and interaction with the araBAD promoter. Cell. 1977 Jul;11(3):545–550. doi: 10.1016/0092-8674(77)90072-1. [DOI] [PubMed] [Google Scholar]
  16. Horwitz A. H., Heffernan L., Cass L., Miyada C. G., Wilcox G. Construction of pBR322-ara hybrid plasmids by in vivo recombination. Mol Gen Genet. 1980;179(3):615–625. doi: 10.1007/BF00271752. [DOI] [PubMed] [Google Scholar]
  17. Lee J. H., Al-Zarban S., Wilcox G. Genetic characterization of the araE gene in Salmonella typhimurium lt2. J Bacteriol. 1981 Apr;146(1):298–304. doi: 10.1128/jb.146.1.298-304.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lee N. L., Gielow W. O., Wallace R. G. Mechanism of araC autoregulation and the domains of two overlapping promoters, Pc and PBAD, in the L-arabinose regulatory region of Escherichia coli. Proc Natl Acad Sci U S A. 1981 Feb;78(2):752–756. doi: 10.1073/pnas.78.2.752. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lee N., Wilcox G., Gielow W., Arnold J., Cleary P., Englesberg E. In vitro activation of the transcription of araBAD operon by araC activator. Proc Natl Acad Sci U S A. 1974 Mar;71(3):634–638. doi: 10.1073/pnas.71.3.634. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Miyada C. G., Horwitz A. H., Cass L. G., Timko J., Wilcox G. DNA sequence of the araC regulatory gene from Escherichia coli B/r. Nucleic Acids Res. 1980 Nov 25;8(22):5267–5274. doi: 10.1093/nar/8.22.5267. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Miyada C. G., Soberón X., Itakura K., Wilcox G. The use of synthetic oligodeoxyribonucleotides to produce specific deletions in the araBAD promoter of Escherichia coli B/r. Gene. 1982 Feb;17(2):167–177. doi: 10.1016/0378-1119(82)90070-1. [DOI] [PubMed] [Google Scholar]
  22. Novick R. P., Clowes R. C., Cohen S. N., Curtiss R., 3rd, Datta N., Falkow S. Uniform nomenclature for bacterial plasmids: a proposal. Bacteriol Rev. 1976 Mar;40(1):168–189. doi: 10.1128/br.40.1.168-189.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. O'Neill M. C., Amass K., de Crombrugghe B. Molecuar model of the DNA interaction site for the cyclic AMP receptor protein. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2213–2217. doi: 10.1073/pnas.78.4.2213. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Ogden S., Haggerty D., Stoner C. M., Kolodrubetz D., Schleif R. The Escherichia coli L-arabinose operon: binding sites of the regulatory proteins and a mechanism of positive and negative regulation. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3346–3350. doi: 10.1073/pnas.77.6.3346. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Queen C., Rosenberg M. A promoter of pBR322 activated by cAMP receptor protein. Nucleic Acids Res. 1981 Jul 24;9(14):3365–3377. doi: 10.1093/nar/9.14.3365. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Smith B. R., Schleif R. Nucleotide sequence of the L-arabinose regulatory region of Escherichia coli K12. J Biol Chem. 1978 Oct 10;253(19):6931–6933. [PubMed] [Google Scholar]
  27. Wallace R. G., Lee N., Fowler A. V. The araC gene of Escherichia coli: transcriptional and translational start-points and complete nucleotide sequence. Gene. 1980 Dec;12(3-4):179–190. doi: 10.1016/0378-1119(80)90100-6. [DOI] [PubMed] [Google Scholar]
  28. Wilcox G., Boulter J., Lee N. Direction of transcription of the regulatory gene araC in Escherichia coli B-r. Proc Natl Acad Sci U S A. 1974 Sep;71(9):3635–3639. doi: 10.1073/pnas.71.9.3635. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES