Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1977 Jan;129(1):305–316. doi: 10.1128/jb.129.1.305-316.1977

Thymidine-requiring mutants of Salmonella typhimurium that are defective in deoxyuridine 5'-phosphate synthesis.

C F Beck, J Neuhard, E Thomassen
PMCID: PMC234928  PMID: 318643

Abstract

In a Salmonella typhimurium strain made diploid for the thy region by introduction of the Escherichia coli episome, F'15, mutants resistant to trimethoprim in the presence of thymidine were selected. One was shown to be defective in deoxyuridine 5'-phosphate (dUMP) synthesis; it requires deoxyuridine or thymidine for growth and is sensitive to trimethoprim in the presence of deoxyuridine. Genetic studies showed that the mutant is mutated in two genes, dcd and dum, located at 70 and 18 min, respectively, on the Salmonella linkage map. The dcd gene cotransduces 95% with udk, the structural gene for uridine kinase. Both mutations are necessary to create a deoxyuridine requirement, providing evidence for the existence of two independent pathways for dUMP synthesis. Pool studies showed that a dum mutation by itself causes a small decrease in the deoxythymidine 5'-triphosphate (dTTP) pool of the cells, whereas a dcd mutation results in a much more marked decrease. The double mutant dcd dum, when incubated in the absence of deoxyuridine, contains barely detectable levels of dTTP. Enzyme analysis revealed that dcd encodes deoxycytidine 5'-triphosphate deaminase. The gene product of the dum gene has not yet been identified; it does not encode either subunit of ribonucleoside diphosphate reductase or deoxyuridine 5'-triphosphate pyrophosphatase. Mutants deleted for the dcd-udk region of the S. typhimurium chromosome were isolated.

Full text

PDF
305

Selected References

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

  1. Beacham I. R., Pritchard R. H. The role of nucleoside phosphorylases in the degradation of deoxyribonucleosides by thymine-requiring mutants of E. coli. Mol Gen Genet. 1971;110(4):289–298. doi: 10.1007/BF00438271. [DOI] [PubMed] [Google Scholar]
  2. Beck C. F., Eisenhardt A. R., Neuhard J. Deoxycytidine triphosphate deaminase of Salmonella typhimurium. Purification and characterization. J Biol Chem. 1975 Jan 25;250(2):609–616. [PubMed] [Google Scholar]
  3. Beck C. F., Ingraham J. L. Location on the chromosome of Salmonella typhimurium of genes governing pyrimidine metabolism. Mol Gen Genet. 1971;111(4):303–316. doi: 10.1007/BF00569782. [DOI] [PubMed] [Google Scholar]
  4. Beck C. F., Ingraham J. L., Neuhard J. Location on the chromosome of Salmonella typhimurium of genes governing pyrimidine metabolism. II. Uridine kinase, cytosine deaminase and thymidine kinase. Mol Gen Genet. 1972;115(3):208–215. doi: 10.1007/BF00268884. [DOI] [PubMed] [Google Scholar]
  5. Beck C. F., Ingraham J. L., Neuhard J., Thomassen E. Metabolism of pyrimidines and pyrimidine nucleosides by Salmonella typhimurium. J Bacteriol. 1972 Apr;110(1):219–228. doi: 10.1128/jb.110.1.219-228.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Beck C. F., Neuhard J., Thomassen E., Ingraham J. L., Kleker E. Salmonella typhimurium mutants defective in cytidine monophosphate kinase (cmk). J Bacteriol. 1974 Dec;120(3):1370–1379. doi: 10.1128/jb.120.3.1370-1379.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Biswas C., Hardy J., Beck W. S. Release of repressor control of ribonucleotide reductase by thymine starvation. J Biol Chem. 1965 Sep;240(9):3631–3640. [PubMed] [Google Scholar]
  8. Charney J. G. NUMERICAL PREDICTION OF CYCLOGENESIS. Proc Natl Acad Sci U S A. 1954 Feb;40(2):99–110. doi: 10.1073/pnas.40.2.99. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Edlin G., Maaloe O. Synthesis and breakdown of messenger RNA without protein synthesis. J Mol Biol. 1966 Feb;15(2):428–434. doi: 10.1016/s0022-2836(66)80118-3. [DOI] [PubMed] [Google Scholar]
  10. Enomoto M., Stocker B. A. Transduction by phage P1kc in Salmonella typhimurium. Virology. 1974 Aug;60(2):503–514. doi: 10.1016/0042-6822(74)90344-4. [DOI] [PubMed] [Google Scholar]
  11. Fuchs J. A., Neuhard J. A mutant of Escherichia coli defective in ribonucleosidediphosphate reductase. 1. Isolation of the mutant as a deoxyuridine auxotroph. Eur J Biochem. 1973 Feb 1;32(3):451–456. doi: 10.1111/j.1432-1033.1973.tb02627.x. [DOI] [PubMed] [Google Scholar]
  12. Guerola N., Ingraham J. L., Cerdá-Olmedo E. Induction of closely linked multiple mutations by nitrosoguanidine. Nat New Biol. 1971 Mar 24;230(12):122–125. doi: 10.1038/newbio230122a0. [DOI] [PubMed] [Google Scholar]
  13. Jensen K. F., Leer J. C., Nygaard P. Thymine utilization in Escherichia coli K12 on the role of deoxyribose 1-phosphate and thymidine phosphorylase. Eur J Biochem. 1973 Dec 17;40(2):345–354. doi: 10.1111/j.1432-1033.1973.tb03203.x. [DOI] [PubMed] [Google Scholar]
  14. Karlström O., Larsson A. Significance of ribonucleotide reduction in the biosynthesis of deoxyribonucleotides in Escherichia coli. Eur J Biochem. 1967 Dec;3(2):164–170. doi: 10.1111/j.1432-1033.1967.tb19512.x. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. MAALOE O., HANAWALT P. C. Thymine deficiency and the normal DNA replication cycle. I. J Mol Biol. 1961 Apr;3:144–155. doi: 10.1016/s0022-2836(61)80041-7. [DOI] [PubMed] [Google Scholar]
  17. Mojica T. Transduction by phage P1CM clr-100 in Salmonella typhimurium. Mol Gen Genet. 1975;138(2):113–126. doi: 10.1007/BF02428116. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Neuhard J. Studies on the acid-soluble nucleotide pool in thymine-requiring mutants of Escherichia coli during thymine starvation. 3. On the regulation of the deoxyadenosine triphosphate and deoxycytidine triphosphate pools of Escherichia coli. Biochim Biophys Acta. 1966 Oct 24;129(1):104–115. doi: 10.1016/0005-2787(66)90012-8. [DOI] [PubMed] [Google Scholar]
  20. Neuhard J., Thomassen E. Altered deoxyribonucleotide pools in P2 eductants of Escherichia coli K-12 due to deletion of the dcd gene. J Bacteriol. 1976 May;126(2):999–1001. doi: 10.1128/jb.126.2.999-1001.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Neuhard J., Thomassen E. Deoxycytidine triphosphate deaminase: identification and function in Salmonella typhimurium. J Bacteriol. 1971 Feb;105(2):657–665. doi: 10.1128/jb.105.2.657-665.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. 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]
  23. O'Donovan G. A., Edlin G., Fuchs J. A., Neuhard J., Thomassen E. Deoxycytidine triphosphate deaminase: characterization of an Escherichia coli mutant deficient in the enzyme. J Bacteriol. 1971 Feb;105(2):666–672. doi: 10.1128/jb.105.2.666-672.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. 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]
  25. OKAZAKI R., KORNBERG A. DEOXYTHYMIDINE KINASE OF ESCHERICHIA COLI. I. PURIFICATION AND SOME PROPERTIES OF THE ENZYME. J Biol Chem. 1964 Jan;239:269–274. [PubMed] [Google Scholar]
  26. Ornellas E. P., Stocker B. A. Relation of lipopolysaccharide character to P1 sensitivity in Salmonella typhimurium. Virology. 1974 Aug;60(2):491–502. doi: 10.1016/0042-6822(74)90343-2. [DOI] [PubMed] [Google Scholar]
  27. Randerath K., Randerath E. Ion-exchange thin-layer chromatography. XV. Preparation, properties and applications of paper-like PEI-cellulose sheets. J Chromatogr. 1966 Apr;22(1):110–117. doi: 10.1016/s0021-9673(01)97076-1. [DOI] [PubMed] [Google Scholar]
  28. STACEY K. A., SIMSON E. IMPROVED METHOD FOR THE ISOLATION OF THYMINE-REQUIRING MUTANTS OF ESCHERICHIA COLI. J Bacteriol. 1965 Aug;90:554–555. doi: 10.1128/jb.90.2.554-555.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Sanderson K. E. Linkage map of Salmonella typhimurium, edition IV. Bacteriol Rev. 1972 Dec;36(4):558–586. doi: 10.1128/br.36.4.558-586.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Sanderson K. E., Ross H., Ziegler L., Mäkelä P. H. F + , Hfr, and F' strains of Salmonella typhimurium and Salmonella abony. Bacteriol Rev. 1972 Dec;36(4):608–637. doi: 10.1128/br.36.4.608-637.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Taylor A. L., Trotter C. D. Linkage map of Escherichia coli strain K-12. Bacteriol Rev. 1972 Dec;36(4):504–524. doi: 10.1128/br.36.4.504-524.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Vosberg H. P., Hoffmann-Berling H. DNA synthesis in nucleotide-permeable Escherichia coli cells. I. Preparation and properties of ether-treated cells. J Mol Biol. 1971 Jun 28;58(3):739–753. doi: 10.1016/0022-2836(71)90037-4. [DOI] [PubMed] [Google Scholar]
  33. Warner H. R. Properties of ribonucleoside diphosphate reductase in nucleotide-permeable cells. J Bacteriol. 1973 Jul;115(1):18–22. doi: 10.1128/jb.115.1.18-22.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Wilkinson R. G., Gemski P., Jr, Stocker B. A. Non-smooth mutants of Salmonella typhimurium: differentiation by phage sensitivity and genetic mapping. J Gen Microbiol. 1972 May;70(3):527–554. doi: 10.1099/00221287-70-3-527. [DOI] [PubMed] [Google Scholar]

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

RESOURCES