Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1982 Jul;151(1):144–152. doi: 10.1128/jb.151.1.144-152.1982

A Regulatory Gene (use) Affecting the Expression of pyrA and Certain Other Pyrimidine Genes

Lee B Bussey 1,, John L Ingraham 1
PMCID: PMC220216  PMID: 7045072

Abstract

The use-1 mutation in Salmonella typhimurium confers a complex and pleiotrophic phenotype which is primarily characterized as a temperature-dependent sensitivity to uracil. This sensitivity can be reversed by arginine or citrulline, but not by ornithine, suggesting that the use-1 mutation affects the synthesis or the activity (or both) of carbamoylphosphate synthetase or ornithine carbamoyltransferase (or both). Activity measurements showed that use-1 caused superrepression of both of these enzymes, especially when uracil was present in the medium. Dihydro-orotase and dihydro-orotate oxidase were also superrepressed, but aspartate carbamoyltransferase and orotate phosphoribosyltransferase were not. Lowered nucleotide triphosphate and guanosine tetra- and pentaphosphate pools in use-1 strains indicated that the mutation affected synthesis or breakdown of all of these phosphorylated compounds, but the UTP pool increased by a larger relative factor in use-1 strains in the presence of uracil. The uracil-sensitive phenotype of the use-1 mutation is a complex response to several environmental factors: temperature, aerobiosis, carbon sources, and uracil concentration. Uracil sensitivity was eliminated by alteration of one or more of these factors. Uracil sensitivity was suppressed by several genetic alterations. These include introduction into use-1 strains of a multi-copy ColE1 derivative which carries the structural gene(s) for carbamoylphosphate synthetase, episomes that carry use, mutations including argR and pyrH, and various unclassified intergenic suppressor mutations. These genetic changes increased significantly the expression of carbamoylphosphate synthetase or ornithine carbamoyltransferase (or both). The activity of use-1 is not known, but the facts that it altered expression of at least four unlinked genes (pyrA, pyrC, pyrD, and argI) and that the Escherichia coli F′133 complemented it establish it as a trans-acting regulatory factor.

Full text

PDF
144

Selected References

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

  1. Abd-el-Al A., Ingraham J. L. Cold sensitivity and other phenotypes resulting from mutation in pyrA gene. J Biol Chem. 1969 Aug 10;244(15):4039–4045. [PubMed] [Google Scholar]
  2. Abd-el-Al A., Ingraham J. L. Control of carbamyl phosphate synthesis in Salmonella typhimurium. J Biol Chem. 1969 Aug 10;244(15):4033–4038. [PubMed] [Google Scholar]
  3. Abdelal A. T., Griego E., Ingraham J. L. Arginine auxotrophic phenotype of mutation in pyrA of Salmonella typhimurium: role of N-acetylornithine in the maturation of mutant carbamylphosphate synthetase. J Bacteriol. 1978 May;134(2):528–536. doi: 10.1128/jb.134.2.528-536.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Abdelal A. T., Ingraham J. L. Carbamylphosphate synthetase from Salmonella typhimurium. Regulations, subunit composition, and function of the subunits. J Biol Chem. 1975 Jun 25;250(12):4410–4417. [PubMed] [Google Scholar]
  5. Abdelal A. T., Kennedy E. H., Nainan O. Ornithine transcarbamylase from Salmonella typhimurium: purification, subunit composition, kinetic analysis, and immunological cross-reactivity. J Bacteriol. 1977 Mar;129(3):1387–1396. doi: 10.1128/jb.129.3.1387-1396.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. BECKWITH J. R., PARDEE A. B., AUSTRIAN R., JACOB F. Coordination of the synthesis of the enzymes in the pyrimidine pathway of E. coli. J Mol Biol. 1962 Dec;5:618–634. doi: 10.1016/s0022-2836(62)80090-4. [DOI] [PubMed] [Google Scholar]
  7. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  8. Cashel M. The control of ribonucleic acid synthesis in Escherichia coli. IV. Relevance of unusual phosphorylated compounds from amino acid-starved stringent strains. J Biol Chem. 1969 Jun 25;244(12):3133–3141. [PubMed] [Google Scholar]
  9. Chan R. K., Botstein D., Watanabe T., Ogata Y. Specialized transduction of tetracycline resistance by phage P22 in Salmonella typhimurium. II. Properties of a high-frequency-transducing lysate. Virology. 1972 Dec;50(3):883–898. doi: 10.1016/0042-6822(72)90442-4. [DOI] [PubMed] [Google Scholar]
  10. Clewell D. B., Helinski D. R. Effect of growth conditions on the formation of the relaxation complex of supercoiled ColE1 deoxyribonucleic acid and protein in Escherichia coli. J Bacteriol. 1972 Jun;110(3):1135–1146. doi: 10.1128/jb.110.3.1135-1146.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Clewell D. B. Nature of Col E 1 plasmid replication in Escherichia coli in the presence of the chloramphenicol. J Bacteriol. 1972 May;110(2):667–676. doi: 10.1128/jb.110.2.667-676.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fyfe J. A., Miller R. L., Krenitsky T. A. Kinetic properties and inhibition of orotidine 5'-phosphate decarboxylase. Effects of some allopurinol metabolites on the enzyme. J Biol Chem. 1973 Jun 10;248(11):3801–3809. [PubMed] [Google Scholar]
  13. GERHART J. C., PARDEE A. B. The enzymology of control by feedback inhibition. J Biol Chem. 1962 Mar;237:891–896. [PubMed] [Google Scholar]
  14. Ingraham J. L., Neuhard J. Cold-sensitive mutants of Salmonella typhimurium defective in uridine monophosphate kinase (pyrH). J Biol Chem. 1972 Oct 10;247(19):6259–6265. [PubMed] [Google Scholar]
  15. Justesen J., Neuhard J. pyrR identical to pyrH in Salmonella typhimurium: control of expression of the pyr genes. J Bacteriol. 1975 Sep;123(3):851–854. doi: 10.1128/jb.123.3.851-854.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Katz L., Helinski D. R. Effect of inhibitors of ribonucleic acid and protein synthesis on the cyclic adenosine monophosphate stimulation of plasmid ColE1 replication. J Bacteriol. 1974 Aug;119(2):450–460. doi: 10.1128/jb.119.2.450-460.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kelln R. A., Kinahan J. J., Foltermann K. F., O'Donovan G. A. Pyrimidine biosynthetic enzymes of Salmonella typhimurium, repressed specifically by growth in the presence of cytidine. J Bacteriol. 1975 Nov;124(2):764–774. doi: 10.1128/jb.124.2.764-774.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kelln R. A., O'Donovan G. A. Isolation and partial characterization of an argR mutant of Salmonella typhimurium. J Bacteriol. 1976 Nov;128(2):528–535. doi: 10.1128/jb.128.2.528-535.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]
  20. Lacroute F., Piérard A., Grenson M., Wiame J. M. The biosynthesis of carbamoyl phosphate in Saccharomyces cerevisiae. J Gen Microbiol. 1965 Jul;40(1):127–142. doi: 10.1099/00221287-40-1-127. [DOI] [PubMed] [Google Scholar]
  21. Lederberg E. M., Cohen S. N. Transformation of Salmonella typhimurium by plasmid deoxyribonucleic acid. J Bacteriol. 1974 Sep;119(3):1072–1074. doi: 10.1128/jb.119.3.1072-1074.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Makoff A. J., Radford A. Genetics and biochemistry of carbamoyl phosphate biosynthesis and its utilization in the pyrimidine biosynthetic pathway. Microbiol Rev. 1978 Jun;42(2):307–328. doi: 10.1128/mr.42.2.307-328.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Mergeay M., Gigot D., Beckmann J., Glansdorff N., Piérard A. Physiology and genetics of carbamoylphosphate synthesis in Escherichia coli K12. Mol Gen Genet. 1974;133(4):299–316. doi: 10.1007/BF00332706. [DOI] [PubMed] [Google Scholar]
  24. Meyers J. A., Sanchez D., Elwell L. P., Falkow S. Simple agarose gel electrophoretic method for the identification and characterization of plasmid deoxyribonucleic acid. J Bacteriol. 1976 Sep;127(3):1529–1537. doi: 10.1128/jb.127.3.1529-1537.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Neidhardt F. C., Bloch P. L., Smith D. F. Culture medium for enterobacteria. J Bacteriol. 1974 Sep;119(3):736–747. doi: 10.1128/jb.119.3.736-747.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Neuhard J. Pyrimidine nucleotide metabolism and pathways of thymidine triphosphate biosynthesis in Salmonella typhimurium. J Bacteriol. 1968 Nov;96(5):1519–1527. doi: 10.1128/jb.96.5.1519-1527.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Neuhard J., Thomassen E. Turnover of the deoxyribonucleoside triphosphates in Escherichia coli 15 T during thymine starvation. Eur J Biochem. 1971 May 11;20(1):36–43. doi: 10.1111/j.1432-1033.1971.tb01359.x. [DOI] [PubMed] [Google Scholar]
  28. O'Donovan G. A., Neuhard J. Pyrimidine metabolism in microorganisms. Bacteriol Rev. 1970 Sep;34(3):278–343. doi: 10.1128/br.34.3.278-343.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Paulus T. J., Switzer R. L. Characterization of pyrimidine-repressible and arginine-repressible carbamyl phosphate synthetases from Bacillus subtilis. J Bacteriol. 1979 Jan;137(1):82–91. doi: 10.1128/jb.137.1.82-91.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. 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]
  31. Porter R. W., Modebe M. O., Stark G. R. Aspartate transcarbamylase. Kinetic studies of the catalytic subunit. J Biol Chem. 1969 Apr 10;244(7):1846–1859. [PubMed] [Google Scholar]
  32. Schwartz M., Neuhard J. Control of expression of the pyr genes in Salmonella typhimurium: effects of variations in uridine and cytidine nucleotide pools. J Bacteriol. 1975 Mar;121(3):814–822. doi: 10.1128/jb.121.3.814-822.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Scott J. F., Roth J. R., Artz S. W. Regulation of histidine operon does not require hisG enzyme. Proc Natl Acad Sci U S A. 1975 Dec;72(12):5021–5025. doi: 10.1073/pnas.72.12.5021. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. VOGEL H. J., BONNER D. M. Acetylornithinase of Escherichia coli: partial purification and some properties. J Biol Chem. 1956 Jan;218(1):97–106. [PubMed] [Google Scholar]

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

RESOURCES