Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1981 Jan;145(1):306–320. doi: 10.1128/jb.145.1.306-320.1981

Recombination of uracil-containing lambda bacteriophages.

J B Hays, B K Duncan, S Boehmer
PMCID: PMC217274  PMID: 6450747

Abstract

Controlled incorporation of uracil into the deoxyribonucleic acid (DNA) of lambda bacteriophages was achieved by growth on dut ung thy mutants of Escherichia coli. The frequency of substitution of uracil for thymine, estimated by alkaline sucrose sedimentation of phage DNA treated in vitro with uracil DNA glycosylase, ranged from 0.17 to 1.9%. The corresponding ratio between the plating efficiencies on wild-type (Ung+) and glycosylase-deficient (Ung-) bacteria ranged from 0.70 to 0.05. If a single-hit dependence of plating efficiency on uracil content is assumed, the probability that any given uracil residue is lethal is approximately 1% (about one-fifth the probability for a pyrimidine dimer). The effect of uracil on recombination was studied in experiments with lambda tandem duplication phages (ethylenediaminetetraacetic acid [EDTA] sensitive), which are converted to single-copy phages (EDTA resistant) by general recombination. For repressed infections (of homoimmune lysogens), recombination was measured by a two-stage assay (DNA extraction, transfection of spheroplasts, and EDTA treatment). The frequencies observed for uracil-containing phages (2 to 4%) were 5 to 10 times higher than control values. However, comparisons with ultraviolet irradiated phages indicated that uracil residues promoted recombination less than 1/100 as efficiently as ultraviolet-induced lesions. Recombination of uracil-containing phages during repressed infections was negligible in recA and partially reduced in recB bacteria. Recombination was very low in ung cells, suggesting that excision repair was responsible for the stimulation. Interestingly, uracil-stimulated recombination was elevated about twofold in xth bacteria.

Full text

PDF
306

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. Chase J. W., Masker W. E. Deoxyribonucleic acid repair in Escherichia coli mutants deficient in the 5'----3' exonuclease activity of deoxyribonucleic acid polymerase I and exonuclease VII. J Bacteriol. 1977 May;130(2):667–675. doi: 10.1128/jb.130.2.667-675.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chase J. W., Richardson C. C. Escherichia coli mutants deficient in exonuclease VII. J Bacteriol. 1977 Feb;129(2):934–947. doi: 10.1128/jb.129.2.934-947.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chase J. W., Richardson C. C. Exonuclease VII of Escherichia coli. Mechanism of action. J Biol Chem. 1974 Jul 25;249(14):4553–4561. [PubMed] [Google Scholar]
  5. 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]
  6. Emmons S. W., MacCosham V., Baldwin R. L. Tandem genetic duplications in phage lambda. III. The frequency of duplication mutants in two derivatives of phage lambda is independent of known recombination systems. J Mol Biol. 1975 Jan 15;91(2):133–146. doi: 10.1016/0022-2836(75)90154-0. [DOI] [PubMed] [Google Scholar]
  7. Emmons S. W., Thomas J. O. Tandem genetic duplications in phage lambda. IV. The locations of spontaneously arising tandem duplications. J Mol Biol. 1975 Jan 15;91(2):147–152. doi: 10.1016/0022-2836(75)90155-2. [DOI] [PubMed] [Google Scholar]
  8. Enquist L. W., Skalka A. Replication of bacteriophage lambda DNA dependent on the function of host and viral genes. I. Interaction of red, gam and rec. J Mol Biol. 1973 Apr 5;75(2):185–212. doi: 10.1016/0022-2836(73)90016-8. [DOI] [PubMed] [Google Scholar]
  9. Gates F. T., Linn S. Endonuclease from Escherichia coli that acts specifically upon duplex DNA damaged by ultraviolet light, osmium tetroxide, acid, or x-rays. J Biol Chem. 1977 May 10;252(9):2802–2807. [PubMed] [Google Scholar]
  10. Gellert M. Formation of covalent circles of lambda DNA by E. coli extracts. Proc Natl Acad Sci U S A. 1967 Jan;57(1):148–155. doi: 10.1073/pnas.57.1.148. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gottesman M. M., Gottesman M. E., Gottesman S., Gellert M. Characterization of bacteriophage lambda reverse as an Escherichia coli phage carrying a unique set of host-derived recombination functions. J Mol Biol. 1974 Sep 15;88(2):471–487. doi: 10.1016/0022-2836(74)90496-3. [DOI] [PubMed] [Google Scholar]
  12. Grossman L., Riazuddin S., Haseltine W. A., Lindan C. Nucleotide excision repair of damaged DNA. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 2):947–955. doi: 10.1101/sqb.1979.043.01.104. [DOI] [PubMed] [Google Scholar]
  13. Guarneros G., Echols H. New mutants of bacteriophage lambda with a specific defect in excision from the host chromosome. J Mol Biol. 1970 Feb 14;47(3):565–574. doi: 10.1016/0022-2836(70)90323-2. [DOI] [PubMed] [Google Scholar]
  14. Hanawalt P. C., Cooper P. K., Ganesan A. K., Smith C. A. DNA repair in bacteria and mammalian cells. Annu Rev Biochem. 1979;48:783–836. doi: 10.1146/annurev.bi.48.070179.004031. [DOI] [PubMed] [Google Scholar]
  15. Haseltine W. A., Gordon L. K., Lindan C. P., Grafstrom R. H., Shaper N. L., Grossman L. Cleavage of pyrimidine dimers in specific DNA sequences by a pyrimidine dimer DNA-glycosylase of M. luteus. Nature. 1980 Jun 26;285(5767):634–641. doi: 10.1038/285634a0. [DOI] [PubMed] [Google Scholar]
  16. Hays J. B., Boehmer S. Antagonists of DNA gyrase inhibit repair and recombination of UV-irradiated phage lambda. Proc Natl Acad Sci U S A. 1978 Sep;75(9):4125–4129. doi: 10.1073/pnas.75.9.4125. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hays J. B., Korba B. E., Konrad E. B. Novel mutations of Escherichia coli that produce recombinogenic lesions in DNA. I. Identification and mapping of arl mutations. J Mol Biol. 1980 May 25;139(3):455–472. doi: 10.1016/0022-2836(80)90141-2. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Howard-Flanders P., Rupp W. D., Wilkins B. M., Cole R. S. DNA replication and recombination after UV irradiation. Cold Spring Harb Symp Quant Biol. 1968;33:195–207. doi: 10.1101/sqb.1968.033.01.023. [DOI] [PubMed] [Google Scholar]
  20. JACOB F., WOLLMAN E. L. Etude génétique d'un bactériophage tempéré d'Escherichia coli. III. Effet du rayonnement ultraviolet sur la recombinaison génétique. Ann Inst Pasteur (Paris) 1955 Jun;88(6):724–749. [PubMed] [Google Scholar]
  21. Kelly R. B., Cozzarelli N. R., Deutscher M. P., Lehman I. R., Kornberg A. Enzymatic synthesis of deoxyribonucleic acid. XXXII. Replication of duplex deoxyribonucleic acid by polymerase at a single strand break. J Biol Chem. 1970 Jan 10;245(1):39–45. [PubMed] [Google Scholar]
  22. 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]
  23. Korba B. E., Hays J. B. Novel mutants of Escherichia coli that produce recombinogenic lesions in DNA. II. Properties of recombinogenic lambda phages grown on bacteria carrying arl mutations. J Mol Biol. 1980 May 25;139(3):473–489. doi: 10.1016/0022-2836(80)90142-4. [DOI] [PubMed] [Google Scholar]
  24. Lin P. F., Howard-Flanders P. Genetic exchanges caused by ultraviolet photoproducts in phage lambda DNA molecules: the role of DNA replication. Mol Gen Genet. 1976 Jul 23;146(2):107–115. doi: 10.1007/BF00268079. [DOI] [PubMed] [Google Scholar]
  25. Lindahl T. DNA glycosylases, endonucleases for apurinic/apyrimidinic sites, and base excision-repair. Prog Nucleic Acid Res Mol Biol. 1979;22:135–192. doi: 10.1016/s0079-6603(08)60800-4. [DOI] [PubMed] [Google Scholar]
  26. Lindahl T., Ljungquist S., Siegert W., Nyberg B., Sperens B. DNA N-glycosidases: properties of uracil-DNA glycosidase from Escherichia coli. J Biol Chem. 1977 May 25;252(10):3286–3294. [PubMed] [Google Scholar]
  27. Lindahl T. New class of enzymes acting on damaged DNA. Nature. 1976 Jan 1;259(5538):64–66. doi: 10.1038/259064a0. [DOI] [PubMed] [Google Scholar]
  28. Ljungquist S. A new endonuclease from Escherichia coli acting at apurinic sites in DNA. J Biol Chem. 1977 May 10;252(9):2808–2814. [PubMed] [Google Scholar]
  29. Ljungquist S., Lindahl T., Howard-Flanders P. Methyl methane sulfonate-sensitive mutant of Escherichia coli deficient in an endonuclease specific for apurinic sites in deoxyribonucleic acid. J Bacteriol. 1976 May;126(2):646–653. doi: 10.1128/jb.126.2.646-653.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. McEntee K. Protein X is the product of the recA gene of Escherichia coli. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5275–5279. doi: 10.1073/pnas.74.12.5275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Milcarek C., Weiss B. Mutants of Escherichia coli with altered deoxyribonucleases. I. Isolation and characterization of mutants for exonuclease 3. J Mol Biol. 1972 Jul 21;68(2):303–318. doi: 10.1016/0022-2836(72)90215-x. [DOI] [PubMed] [Google Scholar]
  32. Nash H. A. Integrative recombination in bacteriophage lambda: analysis of recombinant DNA. J Mol Biol. 1975 Feb 5;91(4):501–514. doi: 10.1016/0022-2836(75)90276-4. [DOI] [PubMed] [Google Scholar]
  33. Parkinson J. S., Huskey R. J. Deletion mutants of bacteriophage lambda. I. Isolation and initial characterization. J Mol Biol. 1971 Mar 14;56(2):369–384. doi: 10.1016/0022-2836(71)90471-2. [DOI] [PubMed] [Google Scholar]
  34. Radman M. An endonuclease from Escherichia coli that introduces single polynucleotide chain scissions in ultraviolet-irradiated DNA. J Biol Chem. 1976 Mar 10;251(5):1438–1445. [PubMed] [Google Scholar]
  35. Signer E. R., Weil J. Recombination in bacteriophage lambda. I. Mutants deficient in general recombination. J Mol Biol. 1968 Jul 14;34(2):261–271. doi: 10.1016/0022-2836(68)90251-9. [DOI] [PubMed] [Google Scholar]
  36. Sinden R. R., Cole R. S. Topography and kinetics of genetic recombination in Escherichia coli treated with psoralen and light. Proc Natl Acad Sci U S A. 1978 May;75(5):2373–2377. doi: 10.1073/pnas.75.5.2373. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. TAMM C., SHAPIRO H. S., LIPSHITZ R., CHARGAFF E. Distribution density of nucleotides within a desoxyribonucleic acid chain. J Biol Chem. 1953 Aug;203(2):673–688. [PubMed] [Google Scholar]
  38. 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]
  39. 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]
  40. 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]
  41. 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]
  42. White B. J., Hochhauser S. J., Cintron N. M., Weiss B. Genetic mapping of xthA, the structural gene for exonuclease III in Escherichia coli K-12. J Bacteriol. 1976 Jun;126(3):1082–1088. doi: 10.1128/jb.126.3.1082-1088.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. 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]

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

RESOURCES