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. 1984 Jul;81(13):4139–4143. doi: 10.1073/pnas.81.13.4139

Multiple regulatory signals in the control region of the Escherichia coli carAB operon.

J Bouvier, J C Patte, P Stragier
PMCID: PMC345384  PMID: 6377309

Abstract

The first reaction in pyrimidine and arginine biosynthesis in Escherichia coli is catalyzed by a single enzyme, carbamoyl-phosphate synthetase (EC 6.3.5.5), the product of the carAB operon. Expression of this operon is cumulatively repressed by arginine and pyrimidines. The nucleotide sequence of the carAB control region was determined and transcriptional starts were localized. Two adjacent promoters, 70 base pairs apart, appear to be used in vivo, the downstream one overlapping a typical arginine operator. The absence of any attenuation-like sequence excludes such a mechanism for pyrimidine-mediated repression. Various fragments of the carA promoter-proximal region were fused in vitro with the lacZ gene. Results obtained with these fusions indicate that (i) translation of the carA gene can be initiated in vivo without an AUG codon but very likely with an UUG or an AUU codon; (ii) the carAB downstream promoter is repressed by arginine; and (iii) the carAB upstream promoter is repressed by pyrimidines and subject to stringent control. When carried by a multicopy plasmid the carAB control region escapes repression by arginine and pyrimidines. The existence of a pyrimidine repressor, present in limiting amounts in the cell, is therefore postulated.

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Selected References

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

  1. Bachmann B. J. Linkage map of Escherichia coli K-12, edition 7. Microbiol Rev. 1983 Jun;47(2):180–230. doi: 10.1128/mr.47.2.180-230.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bukhari A. I., Taylor A. L. Genetic analysis of diaminopimelic acid- and lysine-requiring mutants of Escherichia coli. J Bacteriol. 1971 Mar;105(3):844–854. doi: 10.1128/jb.105.3.844-854.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Casadaban M. J., Martinez-Arias A., Shapira S. K., Chou J. Beta-galactosidase gene fusions for analyzing gene expression in escherichia coli and yeast. Methods Enzymol. 1983;100:293–308. doi: 10.1016/0076-6879(83)00063-4. [DOI] [PubMed] [Google Scholar]
  4. Casadaban M. J. Transposition and fusion of the lac genes to selected promoters in Escherichia coli using bacteriophage lambda and Mu. J Mol Biol. 1976 Jul 5;104(3):541–555. doi: 10.1016/0022-2836(76)90119-4. [DOI] [PubMed] [Google Scholar]
  5. De Felice M., Levinthal M., Iaccarino M., Guardiola J. Growth inhibition as a consequence of antagonism between related amino acids: effect of valine in Escherichia coli K-12. Microbiol Rev. 1979 Mar;43(1):42–58. doi: 10.1128/mr.43.1.42-58.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Débarbouillé M., Raibaud O. Expression of the Escherichia coli malPQ operon remains unaffected after drastic alteration of its promoter. J Bacteriol. 1983 Mar;153(3):1221–1227. doi: 10.1128/jb.153.3.1221-1227.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gigot D., Crabeel M., Feller A., Charlier D., Lissens W., Glansdorff N., Piérard A. Patterns of polarity in the Escherichia coli car AB gene cluster. J Bacteriol. 1980 Aug;143(2):914–920. doi: 10.1128/jb.143.2.914-920.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Glansdorff N., Dambly C., Palchaudhuri S., Crabeel M., Piérard A., Halleux P. Isolation and heteroduplex mapping of a lambda transducing bacteriophage carrying the structural genes for carbamoylphosphate synthase: regulation of enzyme synthesis in Escherichia coli K-12 lysogens. J Bacteriol. 1976 Jul;127(1):302–308. doi: 10.1128/jb.127.1.302-308.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Lissens W., Cunin R., Kelker N., Glansdorff N., Piérard A. In vitro synthesis of Escherichia coli carbamoylphosphate synthase: evidence for participation of the arginine repressor in cumulative repression. J Bacteriol. 1980 Jan;141(1):58–66. doi: 10.1128/jb.141.1.58-66.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Mackie G. A. Cloning of fragments of lambda dapB2 DNA and identification of the dapB gene product. J Biol Chem. 1980 Sep 25;255(18):8928–8935. [PubMed] [Google Scholar]
  11. Mackie G. A., Parsons G. D. Identification of gene products from cloned fragments of the left arm of lambda dapB2. Can J Biochem. 1982 Mar;60(3):338–346. doi: 10.1139/o82-040. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. NOVICK R. P. Micro-iodometric assay for penicillinase. Biochem J. 1962 May;83:236–240. doi: 10.1042/bj0830236. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Navre M., Schachman H. K. Synthesis of aspartate transcarbamoylase in Escherichia coli: transcriptional regulation of the pyrB-pyrI operon. Proc Natl Acad Sci U S A. 1983 Mar;80(5):1207–1211. doi: 10.1073/pnas.80.5.1207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Patte J. C., Morand P., Boy E., Richaud C., Borne F. The relA locus and the regulation of lysine biosynthesis in Escherichia coli. Mol Gen Genet. 1980;179(2):319–325. doi: 10.1007/BF00425459. [DOI] [PubMed] [Google Scholar]
  16. Piette J., Cunin R., Boyen A., Charlier D., Crabeel M., Van Vliet F., Glansdorff N., Squires C., Squires C. L. The regulatory region of the divergent argECBH operon in Escherichia coli K-12. Nucleic Acids Res. 1982 Dec 20;10(24):8031–8048. doi: 10.1093/nar/10.24.8031. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Piette J., Cunin R., Van Vliet F., Charlier D., Crabeel M., Ota Y., Glansdorff N. Homologous control sites and DNA transcription starts in the related argF and argI genes of Escherichia coli K12. EMBO J. 1982;1(7):853–857. doi: 10.1002/j.1460-2075.1982.tb01259.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Piette J., Nyunoya H., Lusty C. J., Cunin R., Weyens G., Crabeel M., Charlier D., Glansdorff N., Piérard A. DNA sequence of the carA gene and the control region of carAB: tandem promoters, respectively controlled by arginine and the pyrimidines, regulate the synthesis of carbamoyl-phosphate synthetase in Escherichia coli K-12. Proc Natl Acad Sci U S A. 1984 Jul;81(13):4134–4138. doi: 10.1073/pnas.81.13.4134. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Piérard A., Glansdorff N., Mergeay M., Wiame J. M. Control of the biosynthesis of carbamoyl phosphate in Escherichia coli. J Mol Biol. 1965 Nov;14(1):23–36. doi: 10.1016/s0022-2836(65)80226-1. [DOI] [PubMed] [Google Scholar]
  20. Piérard A., Lissens W., Halleux P., Cunin R., Glansdorff N. Role of transcriptional regulation and enzyme inactivation in the synthesis of Escherichia coli carbamoylphosphate synthase. J Bacteriol. 1980 Jan;141(1):382–385. doi: 10.1128/jb.141.1.382-385.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Poulis M. I., Shaw D. C., Campbell H. D., Young I. G. In vitro synthesis of the respiratory NADH dehydrogenase of Escherichia coli. Role of UUG as initiation codon. Biochemistry. 1981 Jul 7;20(14):4178–4185. doi: 10.1021/bi00517a035. [DOI] [PubMed] [Google Scholar]
  22. Poulsen P., Jensen K. F., Valentin-Hansen P., Carlsson P., Lundberg L. G. Nucleotide sequence of the Escherichia coli pyrE gene and of the DNA in front of the protein-coding region. Eur J Biochem. 1983 Sep 15;135(2):223–229. doi: 10.1111/j.1432-1033.1983.tb07641.x. [DOI] [PubMed] [Google Scholar]
  23. Richaud F., Richaud C., Haziza C., Patte J. C. Isolement et purification de gènes d'Escherichia coli K12 impliqués dans la biosynthèse de la lysine. C R Seances Acad Sci III. 1981 Nov 9;293(9):507–512. [PubMed] [Google Scholar]
  24. 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]
  25. Sacerdot C., Fayat G., Dessen P., Springer M., Plumbridge J. A., Grunberg-Manago M., Blanquet S. Sequence of a 1.26-kb DNA fragment containing the structural gene for E.coli initiation factor IF3: presence of an AUU initiator codon. EMBO J. 1982;1(3):311–315. doi: 10.1002/j.1460-2075.1982.tb01166.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Sanger F., Coulson A. R. The use of thin acrylamide gels for DNA sequencing. FEBS Lett. 1978 Mar 1;87(1):107–110. doi: 10.1016/0014-5793(78)80145-8. [DOI] [PubMed] [Google Scholar]
  27. Shine J., Dalgarno L. The 3'-terminal sequence of Escherichia coli 16S ribosomal RNA: complementarity to nonsense triplets and ribosome binding sites. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1342–1346. doi: 10.1073/pnas.71.4.1342. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. 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]
  29. Stragier P., Richaud F., Borne F., Patte J. C. Regulation of diaminopimelate decarboxylase synthesis in Escherichia coli. I. Identification of a lysR gene encoding an activator of the lysA gene. J Mol Biol. 1983 Aug 5;168(2):307–320. doi: 10.1016/s0022-2836(83)80020-5. [DOI] [PubMed] [Google Scholar]
  30. Travers A. A. Promoter sequence for stringent control of bacterial ribonucleic acid synthesis. J Bacteriol. 1980 Feb;141(2):973–976. doi: 10.1128/jb.141.2.973-976.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Trotta P. P., Burt M. E., Haschemeyer R. H., Meister A. Reversible dissociation of carbamyl phosphate synthetase into a regulated synthesis subunit and a subunit required for glutamine utilization. Proc Natl Acad Sci U S A. 1971 Oct;68(10):2599–2603. doi: 10.1073/pnas.68.10.2599. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Turnbough C. L., Jr, Hicks K. L., Donahue J. P. Attenuation control of pyrBI operon expression in Escherichia coli K-12. Proc Natl Acad Sci U S A. 1983 Jan;80(2):368–372. doi: 10.1073/pnas.80.2.368. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Turnbough C. L., Jr Regulation of Escherichia coli aspartate transcarbamylase synthesis by guanosine tetraphosphate and pyrimidine ribonucleoside triphosphates. J Bacteriol. 1983 Feb;153(2):998–1007. doi: 10.1128/jb.153.2.998-1007.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Valentin-Hansen P., Aiba H., Schümperli D. The structure of tandem regulatory regions in the deo operon of Escherichia coli K12. EMBO J. 1982;1(3):317–322. doi: 10.1002/j.1460-2075.1982.tb01167.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

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