Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1991 Aug;173(16):5024–5029. doi: 10.1128/jb.173.16.5024-5029.1991

Escherichia coli catabolite gene activator protein mutants defective in positive control of lac operon transcription.

A C Eschenlauer 1, W S Reznikoff 1
PMCID: PMC208191  PMID: 1650341

Abstract

We isolated three Escherichia coli catabolite gene activator protein mutants that are defective in the positive control of transcription initiation from the lac operon promoter region yet retain negative control of transcription from other promoters. One mutant has a substitution of valine for glutamate at residue 72, which lies in the cyclic AMP binding domain and contacts cyclic AMP. The other two mutants have substitutions of asparagine and cysteine for glycine 162, which lies in a surface-exposed turn of the DNA-binding domain. Surprisingly, although all three mutants can repress the lacP2/P3 promoters through the catabolite gene activator protein target site of lac, none displays strong dominance over the ability of wild-type catabolite gene activator protein to stimulate the lacP1 promoter.

Full text

PDF
5029

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. Aiba H., Nakamura T., Mitani H., Mori H. Mutations that alter the allosteric nature of cAMP receptor protein of Escherichia coli. EMBO J. 1985 Dec 1;4(12):3329–3332. doi: 10.1002/j.1460-2075.1985.tb04084.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bell A., Gaston K., Williams R., Chapman K., Kolb A., Buc H., Minchin S., Williams J., Busby S. Mutations that alter the ability of the Escherichia coli cyclic AMP receptor protein to activate transcription. Nucleic Acids Res. 1990 Dec 25;18(24):7243–7250. doi: 10.1093/nar/18.24.7243. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bracco L., Kotlarz D., Kolb A., Diekmann S., Buc H. Synthetic curved DNA sequences can act as transcriptional activators in Escherichia coli. EMBO J. 1989 Dec 20;8(13):4289–4296. doi: 10.1002/j.1460-2075.1989.tb08615.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brandenburger A., Godson G. N., Radman M., Glickman B. W., van Sluis C. A., Doubleday O. P. Radiation-induced base substitution mutagenesis in single-stranded DNA phage M13. Nature. 1981 Nov 12;294(5837):180–182. doi: 10.1038/294180a0. [DOI] [PubMed] [Google Scholar]
  6. Donnelly C. E., Reznikoff W. S. Mutations in the lac P2 promoter. J Bacteriol. 1987 May;169(5):1812–1817. doi: 10.1128/jb.169.5.1812-1817.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Garges S., Adhya S. Cyclic AMP-induced conformational change of cyclic AMP receptor protein (CRP): intragenic suppressors of cyclic AMP-independent CRP mutations. J Bacteriol. 1988 Apr;170(4):1417–1422. doi: 10.1128/jb.170.4.1417-1422.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Herbert M., Kolb A., Buc H. Overlapping promoters and their control in Escherichia coli: the gal case. Proc Natl Acad Sci U S A. 1986 May;83(9):2807–2811. doi: 10.1073/pnas.83.9.2807. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Irwin N., Ptashne M. Mutants of the catabolite activator protein of Escherichia coli that are specifically deficient in the gene-activation function. Proc Natl Acad Sci U S A. 1987 Dec;84(23):8315–8319. doi: 10.1073/pnas.84.23.8315. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kenyon C. J., Brent R., Ptashne M., Walker G. C. Regulation of damage-inducible genes in Escherichia coli. J Mol Biol. 1982 Sep 25;160(3):445–457. doi: 10.1016/0022-2836(82)90307-2. [DOI] [PubMed] [Google Scholar]
  11. Li X. M., Krakow J. S. Characterization of nine monoclonal antibodies against the Escherichia coli cyclic AMP receptor protein. J Biol Chem. 1985 Apr 10;260(7):4378–4383. [PubMed] [Google Scholar]
  12. Malan T. P., Kolb A., Buc H., McClure W. R. Mechanism of CRP-cAMP activation of lac operon transcription initiation activation of the P1 promoter. J Mol Biol. 1984 Dec 25;180(4):881–909. doi: 10.1016/0022-2836(84)90262-6. [DOI] [PubMed] [Google Scholar]
  13. Menendez M., Kolb A., Buc H. A new target for CRP action at the malT promoter. EMBO J. 1987 Dec 20;6(13):4227–4234. doi: 10.1002/j.1460-2075.1987.tb02771.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Musso R. E., Di Lauro R., Adhya S., de Crombrugghe B. Dual control for transcription of the galactose operon by cyclic AMP and its receptor protein at two interspersed promoters. Cell. 1977 Nov;12(3):847–854. doi: 10.1016/0092-8674(77)90283-5. [DOI] [PubMed] [Google Scholar]
  15. Okamoto K., Freundlich M. Mechanism for the autogenous control of the crp operon: transcriptional inhibition by a divergent RNA transcript. Proc Natl Acad Sci U S A. 1986 Jul;83(14):5000–5004. doi: 10.1073/pnas.83.14.5000. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Plaskon R. R., Wartell R. M. Sequence distributions associated with DNA curvature are found upstream of strong E. coli promoters. Nucleic Acids Res. 1987 Jan 26;15(2):785–796. doi: 10.1093/nar/15.2.785. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Ren Y. L., Garges S., Adhya S., Krakow J. S. Cooperative DNA binding of heterologous proteins: evidence for contact between the cyclic AMP receptor protein and RNA polymerase. Proc Natl Acad Sci U S A. 1988 Jun;85(12):4138–4142. doi: 10.1073/pnas.85.12.4138. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Sheen J. Y., Seed B. Electrolyte gradient gels for DNA sequencing. Biotechniques. 1988 Nov-Dec;6(10):942–944. [PubMed] [Google Scholar]
  19. Singer M., Baker T. A., Schnitzler G., Deischel S. M., Goel M., Dove W., Jaacks K. J., Grossman A. D., Erickson J. W., Gross C. A. A collection of strains containing genetically linked alternating antibiotic resistance elements for genetic mapping of Escherichia coli. Microbiol Rev. 1989 Mar;53(1):1–24. doi: 10.1128/mr.53.1.1-24.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Straney D. C., Straney S. B., Crothers D. M. Synergy between Escherichia coli CAP protein and RNA polymerase in the lac promoter open complex. J Mol Biol. 1989 Mar 5;206(1):41–57. doi: 10.1016/0022-2836(89)90522-6. [DOI] [PubMed] [Google Scholar]
  21. Warwicker J., Engelman B. P., Steitz T. A. Electrostatic calculations and model-building suggest that DNA bound to CAP is sharply bent. Proteins. 1987;2(4):283–289. doi: 10.1002/prot.340020404. [DOI] [PubMed] [Google Scholar]
  22. Weber I. T., Steitz T. A. Structure of a complex of catabolite gene activator protein and cyclic AMP refined at 2.5 A resolution. J Mol Biol. 1987 Nov 20;198(2):311–326. doi: 10.1016/0022-2836(87)90315-9. [DOI] [PubMed] [Google Scholar]
  23. Wu H. M., Crothers D. M. The locus of sequence-directed and protein-induced DNA bending. Nature. 1984 Apr 5;308(5959):509–513. doi: 10.1038/308509a0. [DOI] [PubMed] [Google Scholar]
  24. Xiong X. F., de la Cruz N., Reznikoff W. S. Downstream deletion analysis of the lac promoter. J Bacteriol. 1991 Aug;173(15):4570–4577. doi: 10.1128/jb.173.15.4570-4577.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. de Crombrugghe B., Busby S., Buc H. Cyclic AMP receptor protein: role in transcription activation. Science. 1984 May 25;224(4651):831–838. doi: 10.1126/science.6372090. [DOI] [PubMed] [Google Scholar]

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

RESOURCES