Abstract
A new assay specific for uracil-DNA glycosylase is described, Escherichia coli mutants partially and totally deficient in uracil-DNA glycosylase activity have been isolated by using this assay in mass-screening procedures. These have been designated ung mutants. The ung gene maps between tyrA and nadB on the E. coli chromosome. T4 phage containing uracil in their DNA grow on the most glycosylase-deficient hosts but are unable to grow on wild-type bacteria. This provides a simple spot test for the ung genotype. The ung mutants show slightly higher rates of spontaneous mutation to antibiotic resistance. Taken together, these results suggest a central role for uracil-DNA glycosylase in the initiation of an excision repair pathway for the exclusion of uracil from DNA.
Full text
PDFImages in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Bachmann B. J., Low K. B., Taylor A. L. Recalibrated linkage map of Escherichia coli K-12. Bacteriol Rev. 1976 Mar;40(1):116–167. doi: 10.1128/br.40.1.116-167.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Cone R., Duncan J., Hamilton L., Friedberg E. C. Partial purification and characterization of a uracil DNA N-glycosidase from Bacillus subtilis. Biochemistry. 1977 Jul 12;16(14):3194–3201. doi: 10.1021/bi00633a024. [DOI] [PubMed] [Google Scholar]
- 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]
- Da Roza R., Friedberg E. C., Duncan B. K., Warner H. R. Repair of nitrous acid damage to DNA in Escherichia coli. Biochemistry. 1977 Nov 1;16(22):4934–4939. doi: 10.1021/bi00641a030. [DOI] [PubMed] [Google Scholar]
- Duncan B. K., Warner H. R. Metabolism of uracil-containing DNA: degradation of bacteriophage PBS2 DNA in Bacillus subtilis. J Virol. 1977 Jun;22(3):835–838. doi: 10.1128/jvi.22.3.835-838.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Duncan J., Hamilton L., Friedberg E. C. Enzymatic degradation of uracil-containing DNA. II. Evidence for N-glycosidase and nuclease activities in unfractionated extracts of Bacillus subtilis. J Virol. 1976 Aug;19(2):338–345. doi: 10.1128/jvi.19.2.338-345.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Friedberg E. C., Ganesan A. K., Minton K. N-Glycosidase activity in extracts of Bacillus subtilis and its inhibition after infection with bacteriophage PBS2. J Virol. 1975 Aug;16(2):315–321. doi: 10.1128/jvi.16.2.315-321.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gates F. T., 3rd, Linn S. Endonuclease V of Escherichia coli. J Biol Chem. 1977 Mar 10;252(5):1647–1653. [PubMed] [Google Scholar]
- Isolation and characterization of a Bacillus subtilis mutant with a defective N-glycosidase activity for uracil-containing deoxyribonucleic acid. J Bacteriol. 1977 Aug;131(2):438–445. doi: 10.1128/jb.131.2.438-445.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lindahl T. An N-glycosidase from Escherichia coli that releases free uracil from DNA containing deaminated cytosine residues. Proc Natl Acad Sci U S A. 1974 Sep;71(9):3649–3653. doi: 10.1073/pnas.71.9.3649. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- Lindahl T., Nyberg B. Heat-induced deamination of cytosine residues in deoxyribonucleic acid. Biochemistry. 1974 Jul 30;13(16):3405–3410. doi: 10.1021/bi00713a035. [DOI] [PubMed] [Google Scholar]
- 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]
- Raetz C. R., Larson T. J., Dowhan W. Gene cloning for the isolation of enzymes of membrane lipid synthesis: phosphatidylserine synthase overproduction in Escherichia coli. Proc Natl Acad Sci U S A. 1977 Apr;74(4):1412–1416. doi: 10.1073/pnas.74.4.1412. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Snyder L., Gold L., Kutter E. A gene of bacteriophage T4 whose product prevents true late transcription on cytosine-containing T4 DNA. Proc Natl Acad Sci U S A. 1976 Sep;73(9):3098–3102. doi: 10.1073/pnas.73.9.3098. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tritz G. J., Matney T. S., Gholson R. K. Mapping of the nadB locus adjacent to a previously undescribed purine locus in Escherichia coli K-12. J Bacteriol. 1970 May;102(2):377–381. doi: 10.1128/jb.102.2.377-381.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- 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]
- Weiss B., Milcarek C. Mass screening for mutants with altered DNases by microassay techniques. Methods Enzymol. 1974;29:180–193. doi: 10.1016/0076-6879(74)29020-7. [DOI] [PubMed] [Google Scholar]