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. 1981 Feb;145(2):687–695. doi: 10.1128/jb.145.2.687-695.1981

Synthesis and metabolism of uracil-containing deoxyribonucleic acid in Escherichia coli.

H R Warner, B K Duncan, C Garrett, J Neuhard
PMCID: PMC217167  PMID: 6109711

Abstract

Significant amounts of uracil were found in the deoxyribonucleic acids (DNAs) of Escherichia coli mutants deficient in both uracil-DNA glycosylase (ung) and deoxyuridine 5'-triphosphate nucleotidohydrolase (dut) activities, whereas little uracil was found in the DNAs of wild-type cells and cells deficient in only one of these two activities. The amounts of uracil found in the DNAs of dut ung mutants were directly related to the growth temperature of the cultures, apparently because the deoxyuridine 5'-triphosphate nucleotidohydrolase synthesized by dut mutants was temperature sensitive. The dut mutant used failed to grow exponentially, became filamentous at temperatures above 25 degrees C, and exhibited a hyperrec phenotype; however, the ung mutation suppressed all of these effects. Although the dut ung mutants grew exponentially at all temperatures, their growth rates were always slower than the growth rate of the wild type. Since pool size measurements indicated that both deoxyuridine triphosphate and deoxythymidine triphosphate pools were markedly elevated in dut mutants, the reduced growth rate of dut ung cells apparently was due to the actual presence of uracil in the DNA, rather than to a deficiency of deoxyuridine triphosphate and deoxyribosylthymine triphosphate for DNA synthesis. The presence of uracil in E. coli donor DNA also markedly reduced the recombination frequency when the recipient cells were ung+, indicating that DNA repair commenced before the entering DNA could be replicated.

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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. Pedigrees of some mutant strains of Escherichia coli K-12. Bacteriol Rev. 1972 Dec;36(4):525–557. doi: 10.1128/br.36.4.525-557.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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]
  3. Duncan B. K., Rockstroh P. A., Warner H. R. Escherichia coli K-12 mutants deficient in uracil-DNA glycosylase. J Bacteriol. 1978 Jun;134(3):1039–1045. doi: 10.1128/jb.134.3.1039-1045.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. FRASER D., JERREL E. A. The amino acid composition of T3 bacteriophage. J Biol Chem. 1953 Nov;205(1):291–295. [PubMed] [Google Scholar]
  6. Fisher E. F., Caruthers M. H. Studies on gene control regions XII. The functional significance of a lac operator constitutive mutation. Nucleic Acids Res. 1979 Sep 25;7(2):401–416. doi: 10.1093/nar/7.2.401. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gates F. T., 3rd, Linn S. Endonuclease V of Escherichia coli. J Biol Chem. 1977 Mar 10;252(5):1647–1653. [PubMed] [Google Scholar]
  8. Goeddel D. V., Yansura D. G., Caruthers M. H. How lac repressor recognizes lac operator. Proc Natl Acad Sci U S A. 1978 Aug;75(8):3578–3582. doi: 10.1073/pnas.75.8.3578. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hochhauser S. J., Weiss B. Escherichia coli mutants deficient in deoxyuridine triphosphatase. J Bacteriol. 1978 Apr;134(1):157–166. doi: 10.1128/jb.134.1.157-166.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hoess R. H., Herman R. K. Isolation and characterization of mutator strains of Escherichia coli K-12. J Bacteriol. 1975 May;122(2):474–484. doi: 10.1128/jb.122.2.474-484.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Konrad E. B., Lehman I. R. Novel mutants of Escherichia coli that accumulate very small DNA replicative intermediates. Proc Natl Acad Sci U S A. 1975 Jun;72(6):2150–2154. doi: 10.1073/pnas.72.6.2150. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Konrad E. B. Method for the isolation of Escherichia coli mutants with enhanced recombination between chromosomal duplications. J Bacteriol. 1977 Apr;130(1):167–172. doi: 10.1128/jb.130.1.167-172.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Neuhard J., Maltman K. L., Warren R. A. Bacteriophage phi W-14-infected Pseudomonas acidovorans synthesizes hydroxymethyldeoxyuridine triphosphate. J Virol. 1980 May;34(2):347–353. doi: 10.1128/jvi.34.2.347-353.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Olivera B. M., Manlapaz-Ramos P., Warner H. R., Duncan B. K. DNA intermediates at the Escherichia coli replication fork. II. Studies using dut and ung mutants in vitro. J Mol Biol. 1979 Mar 5;128(3):265–275. doi: 10.1016/0022-2836(79)90087-1. [DOI] [PubMed] [Google Scholar]
  15. Riazuddin S., Lindahl T. Properties of 3-methyladenine-DNA glycosylase from Escherichia coli. Biochemistry. 1978 May 30;17(11):2110–2118. doi: 10.1021/bi00604a014. [DOI] [PubMed] [Google Scholar]
  16. Shlomai J., Kornberg A. Deoxyuridine triphosphatase of Escherichia coli. Purification, properties, and use as a reagent to reduce uracil incorporation into DNA. J Biol Chem. 1978 May 10;253(9):3305–3312. [PubMed] [Google Scholar]
  17. Tye B. K., Chien J., Lehman I. R., Duncan B. K., Warner H. R. Uracil incorporation: a source of pulse-labeled DNA fragments in the replication of the Escherichia coli chromosome. Proc Natl Acad Sci U S A. 1978 Jan;75(1):233–237. doi: 10.1073/pnas.75.1.233. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Tye B. K., Lehman I. R. Excision repair of uracil incorporated in DNA as a result of a defect in dUTPase. J Mol Biol. 1977 Dec 5;117(2):293–306. doi: 10.1016/0022-2836(77)90128-0. [DOI] [PubMed] [Google Scholar]
  19. Tye B. K., Nyman P. O., Lehman I. R., Hochhauser S., Weiss B. Transient accumulation of Okazaki fragments as a result of uracil incorporation into nascent DNA. Proc Natl Acad Sci U S A. 1977 Jan;74(1):154–157. doi: 10.1073/pnas.74.1.154. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Warner H. R., Demple B. F., Deutsch W. A., Kane C. M., Linn S. Apurinic/apyrimidinic endonucleases in repair of pyrimidine dimers and other lesions in DNA. Proc Natl Acad Sci U S A. 1980 Aug;77(8):4602–4606. doi: 10.1073/pnas.77.8.4602. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Warner H. R., Duncan B. K. In vivo synthesis and properties of uracil-containing DNA. Nature. 1978 Mar 2;272(5648):32–34. doi: 10.1038/272032a0. [DOI] [PubMed] [Google Scholar]
  22. Warner H. R., Thompson R. B., Mozer T. J., Duncan B. K. The properties of a bacteriophage T5 mutant unable to induce deoxyuridine 5'-triphosphate nucleotidohydrolase. Synthesis of uracil-containing T5 deoxyribonucleic acid. J Biol Chem. 1979 Aug 25;254(16):7534–7539. [PubMed] [Google Scholar]
  23. Zieg J., Maples V. F., Kushner S. R. Recombinant levels of Escherichia coli K-12 mutants deficient in various replication, recombination, or repair genes. J Bacteriol. 1978 Jun;134(3):958–966. doi: 10.1128/jb.134.3.958-966.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]

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