Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1984 Jun;158(3):948–953. doi: 10.1128/jb.158.3.948-953.1984

Promoter mutation causing catabolite repression of the Salmonella typhimurium leucine operon.

R M Gemmill, M Tripp, S B Friedman, J M Calvo
PMCID: PMC215533  PMID: 6327652

Abstract

Two mutations that affect expression of the Salmonella typhimurium leu operon were investigated. leu operon DNA from these mutant strains was cloned, and nucleotide sequences of the leu control regions were determined. leu-500, which eliminates expression of all four leu genes simultaneously, is a point mutation in the -10 region of the leu promoter. leu-2012 is a point mutation within the -35 region of the leu promoter. leu-2012 suppressed leucine auxotrophy caused by leu-500 only when the medium contained a carbon source that does not cause catabolite repression. A cya mutation (adenylate cyclase deficiency) introduced into the leu-500 leu-2012 strain caused leu enzymes to be made only if cAMP was supplied exogenously. A leu-500 leu-2012 strain containing a crp mutation (cAMP receptor protein deficiency), on the other hand, could not make leu enzymes even in the presence of cAMP. In vitro transcription experiments demonstrated that the leu-2012 mutation created a new transcription initiation site. RNA polymerase utilized this site in vitro in the absence of added cAMP receptor protein and cAMP.

Full text

PDF
948

Images in this article

Selected References

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

  1. Alper M. D., Ames B. N. Transport of antibiotics and metabolite analogs by systems under cyclic AMP control: positive selection of Salmonella typhimurium cya and crp mutants. J Bacteriol. 1978 Jan;133(1):149–157. doi: 10.1128/jb.133.1.149-157.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. BURNS R. O., UMBARGER H. E., GROSS S. R. THE BIOSYNTHESIS OF LEUCINE. III. THE CONVERSION OF ALPHA-HYDROXY-BETA-CARBOXYISOCAPROATE TO ALPHA-KETOISOCAPROATE. Biochemistry. 1963 Sep-Oct;2:1053–1058. doi: 10.1021/bi00905a024. [DOI] [PubMed] [Google Scholar]
  3. DAVIS B. D., MINGIOLI E. S. Mutants of Escherichia coli requiring methionine or vitamin B12. J Bacteriol. 1950 Jul;60(1):17–28. doi: 10.1128/jb.60.1.17-28.1950. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dallas W. S., Tseng Y., Dobrogosz W. J. Regulation of membrane functions and fatty acid composition in Escherichia coli by cyclic AMP receptor protein. Arch Biochem Biophys. 1976 Jul;175(1):295–302. doi: 10.1016/0003-9861(76)90511-7. [DOI] [PubMed] [Google Scholar]
  5. Davis M. G., Calvo J. M. Relationship between messenger ribonucleic acid and enzyme levels specified by the leucine operon of Escherichia coli K-12. J Bacteriol. 1977 Sep;131(3):997–1007. doi: 10.1128/jb.131.3.997-1007.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dubnau E., Margolin P. Suppression of promoter mutations by the pleiotropic supx mutations. Mol Gen Genet. 1972;117(2):91–112. doi: 10.1007/BF00267607. [DOI] [PubMed] [Google Scholar]
  7. Friedman S. B., Margolin P. Evidence for an altered operator specificity: catabolite repression control of the leucine operon in Salmonella typhimurium. J Bacteriol. 1968 Jun;95(6):2263–2269. doi: 10.1128/jb.95.6.2263-2269.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gemmill R. M., Jones J. W., Haughn G. W., Calvo J. M. Transcription initiation sites of the leucine operons of Salmonella typhimurium and Escherichia coli. J Mol Biol. 1983 Oct 15;170(1):39–59. doi: 10.1016/s0022-2836(83)80226-5. [DOI] [PubMed] [Google Scholar]
  9. Gemmill R. M., Wessler S. R., Keller E. B., Calvo J. M. leu operon of Salmonella typhimurium is controlled by an attenuation mechanism. Proc Natl Acad Sci U S A. 1979 Oct;76(10):4941–4945. doi: 10.1073/pnas.76.10.4941. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Graf L. H., Jr, Burns R. O. The supX-leu-500 mutations and expression of the leucine operon. Mol Gen Genet. 1973 Nov 22;126(4):291–301. doi: 10.1007/BF00269439. [DOI] [PubMed] [Google Scholar]
  11. Hertzberg K. M., Gemmill R., Jones J., Calvo J. M. Cloning of an EcoRI-generated fragment of the leucine operon of Salmonella typhimurium. Gene. 1980 Jan;8(2):135–152. doi: 10.1016/0378-1119(80)90033-5. [DOI] [PubMed] [Google Scholar]
  12. Kupersztoch-Portnoy Y. M., Lovett M. A., Helinski D. R. Strand and site specificity of the relaxation event for the relaxation complex of the antibiotic resistance plasmid R6K. Biochemistry. 1974 Dec 31;13(27):5484–5490. doi: 10.1021/bi00724a005. [DOI] [PubMed] [Google Scholar]
  13. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  14. Le Grice S. F., Matzura H., Marcoli R., Iida S., Bickle T. A. The catabolite-sensitive promoter for the chloramphenicol acetyl transferase gene is preceded by two binding sites for the catabolite gene activator protein. J Bacteriol. 1982 Apr;150(1):312–318. doi: 10.1128/jb.150.1.312-318.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lee F., Yanofsky C. Transcription termination at the trp operon attenuators of Escherichia coli and Salmonella typhimurium: RNA secondary structure and regulation of termination. Proc Natl Acad Sci U S A. 1977 Oct;74(10):4365–4369. doi: 10.1073/pnas.74.10.4365. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Luginbuhl G. H., Hofler J. G., Decedue C. J., Burns R. O. Biodegradative L-threonine deaminase of Salmonella typhimurium. J Bacteriol. 1974 Oct;120(1):559–561. doi: 10.1128/jb.120.1.559-561.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. MARGOLIN P. Genetic fine structure of the leucine operon in Salmonella. Genetics. 1963 Mar;48:441–457. doi: 10.1093/genetics/48.3.441. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. 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]
  20. 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]
  21. Rosenberg M., Court D. Regulatory sequences involved in the promotion and termination of RNA transcription. Annu Rev Genet. 1979;13:319–353. doi: 10.1146/annurev.ge.13.120179.001535. [DOI] [PubMed] [Google Scholar]
  22. Searles L. L., Wessler S. R., Calvo J. M. Transcription attenuation is the major mechanism by which the leu operon of Salmonella typhimurium is controlled. J Mol Biol. 1983 Jan 25;163(3):377–394. doi: 10.1016/0022-2836(83)90064-5. [DOI] [PubMed] [Google Scholar]
  23. Siebenlist U. RNA polymerase unwinds an 11-base pair segment of a phage T7 promoter. Nature. 1979 Jun 14;279(5714):651–652. doi: 10.1038/279651a0. [DOI] [PubMed] [Google Scholar]
  24. Simpson R. B. Interaction of the cAMP receptor protein with the lac promoter. Nucleic Acids Res. 1980 Feb 25;8(4):759–766. [PMC free article] [PubMed] [Google Scholar]
  25. Smith D. R., Calvo J. M. Nucleotide sequence of the E coli gene coding for dihydrofolate reductase. Nucleic Acids Res. 1980 May 24;8(10):2255–2274. doi: 10.1093/nar/8.10.2255. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Sternglanz R., DiNardo S., Voelkel K. A., Nishimura Y., Hirota Y., Becherer K., Zumstein L., Wang J. C. Mutations in the gene coding for Escherichia coli DNA topoisomerase I affect transcription and transposition. Proc Natl Acad Sci U S A. 1981 May;78(5):2747–2751. doi: 10.1073/pnas.78.5.2747. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Taniguchi T., O'Neill M., de Crombrugghe B. Interaction site of Escherichia coli cyclic AMP receptor protein on DNA of galactose operon promoters. Proc Natl Acad Sci U S A. 1979 Oct;76(10):5090–5094. doi: 10.1073/pnas.76.10.5090. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Trucksis M., Depew R. E. Identification and localization of a gene that specifies production of Escherichia coli DNA topoisomerase I. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2164–2168. doi: 10.1073/pnas.78.4.2164. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Ullmann A., Joseph E., Danchin A. Cyclic AMP as a modulator of polarity in polycistronic transcriptional units. Proc Natl Acad Sci U S A. 1979 Jul;76(7):3194–3197. doi: 10.1073/pnas.76.7.3194. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Youderian P., Bouvier S., Susskind M. M. Sequence determinants of promoter activity. Cell. 1982 Oct;30(3):843–853. doi: 10.1016/0092-8674(82)90289-6. [DOI] [PubMed] [Google Scholar]

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

RESOURCES