Abstract
An activity has been purified 350-fold from extracts of mouse plasmacytoma cells that forms 5-hydroxymethyluracil (alpha-hydroxythymine) and apyrimidinic sites with phage SPO1 DNA, which contains this base in place of thymine. This DNA glycosylase presumably functions to eliminate hydroxymethyluracil, a major thymine-derived DNA lesion produced by ionizing radiation and oxidative damage. The enzyme has no cofactor requirement and is active in EDTA. Neither intermediate formation nor hydrolysis of hydroxymethyl-deoxyuridine or hydroxymethyldeoxyuridine monophosphate was detected. The enzyme does not cleave apyrimidinic sites in DNA. It does release uracil from the uracil-containing DNA of phage PBS2, but this activity is less than 2% of the predominant uracil DNA glycosylase activity of the cell, which is separated by phosphocellulose chromatography. The major uracil DNA glycosylase does not release hydroxymethyluracil from SPO1 DNA. The hydroxymethyluracil glycosylase is also separated upon phosphocelluose chromatography from a thymine glycol DNA glycosylase activity that is accompanied by an apyrimidinic endonuclease activity.
Full text
PDF![4003](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fb81/345356/1174535f9e7d/pnas00614-0081.png)
![4004](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fb81/345356/fc9cb398b249/pnas00614-0082.png)
![4005](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fb81/345356/522c52598e5d/pnas00614-0083.png)
![4006](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fb81/345356/da3439ef96af/pnas00614-0084.png)
![4007](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fb81/345356/25e51131609d/pnas00614-0085.png)
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Ames B. N. Dietary carcinogens and anticarcinogens. Oxygen radicals and degenerative diseases. Science. 1983 Sep 23;221(4617):1256–1264. doi: 10.1126/science.6351251. [DOI] [PubMed] [Google Scholar]
- Anagnostopoulos C., Spizizen J. REQUIREMENTS FOR TRANSFORMATION IN BACILLUS SUBTILIS. J Bacteriol. 1961 May;81(5):741–746. doi: 10.1128/jb.81.5.741-746.1961. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Anderson C. T., Friedberg E. C. The presence of nuclear and mitochondrial uracil-DNA glycosylase in extracts of human KB cells. Nucleic Acids Res. 1980 Feb 25;8(4):875–888. [PMC free article] [PubMed] [Google Scholar]
- Armel P. R., Strniste G. F., Wallace S. S. Studies on Escherichia coli x-ray endonuclease specificity. Roles of hydroxyl and reducing radicals in the production of DNA lesions. Radiat Res. 1977 Feb;69(2):328–338. [PubMed] [Google Scholar]
- Bacchetti S., Benne R. Purification and characterization of an endonuclease from calf thymus acting on irradiated DNA. Biochim Biophys Acta. 1975 May 16;390(3):285–297. doi: 10.1016/0005-2787(75)90349-4. [DOI] [PubMed] [Google Scholar]
- 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: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
- Breimer L. H., Lindahl T. DNA glycosylase activities for thymine residues damaged by ring saturation, fragmentation, or ring contraction are functions of endonuclease III in Escherichia coli. J Biol Chem. 1984 May 10;259(9):5543–5548. [PubMed] [Google Scholar]
- Breimer L. H. Urea--DNA glycosylase in mammalian cells. Biochemistry. 1983 Aug 30;22(18):4192–4197. doi: 10.1021/bi00287a005. [DOI] [PubMed] [Google Scholar]
- Breimer L., Lindahl T. A DNA glycosylase from Escherichia coli that releases free urea from a polydeoxyribonucleotide containing fragments of base residues. Nucleic Acids Res. 1980 Dec 20;8(24):6199–6211. doi: 10.1093/nar/8.24.6199. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Brent T. P. Characterization of human enzymes specific for damaged DNA: resolution of endonuclease for irradiated DNA from an apparent N-glycosidase active on alkylated DNA. Nucleic Acids Res. 1977 Jul;4(7):2445–2454. doi: 10.1093/nar/4.7.2445. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chetsanga C. J., Lindahl T. Release of 7-methylguanine residues whose imidazole rings have been opened from damaged DNA by a DNA glycosylase from Escherichia coli. Nucleic Acids Res. 1979 Aug 10;6(11):3673–3684. doi: 10.1093/nar/6.11.3673. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cregg J. M., Stewart C. R. Terminal redundancy of "high frequency of recombination" markers of Bacillus subtilis phage SPO1. Virology. 1978 May 15;86(2):530–541. doi: 10.1016/0042-6822(78)90091-0. [DOI] [PubMed] [Google Scholar]
- Demple B., Gates F. T., 3rd, Linn S. Purification and properties of Escherichia coli endodeoxyribonuclease V. Methods Enzymol. 1980;65(1):224–231. doi: 10.1016/s0076-6879(80)65031-9. [DOI] [PubMed] [Google Scholar]
- Demple B., Linn S. DNA N-glycosylases and UV repair. Nature. 1980 Sep 18;287(5779):203–208. doi: 10.1038/287203a0. [DOI] [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]
- 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]
- Helland D., Nes I. F., Kleppe K. Mammalian DNA-repair endonuclease acts only on supercoiled DNA. FEBS Lett. 1982 Jun 1;142(1):121–124. doi: 10.1016/0014-5793(82)80233-0. [DOI] [PubMed] [Google Scholar]
- Hemphill H. E., Whiteley H. R. Bacteriophages of Bacillus subtilis. Bacteriol Rev. 1975 Sep;39(3):257–315. doi: 10.1128/br.39.3.257-315.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Iida S., Hayatsu H. The permanganate oxidation of thymine. Biochim Biophys Acta. 1970 Jul 16;213(1):1–13. doi: 10.1016/0005-2787(70)90002-x. [DOI] [PubMed] [Google Scholar]
- Kane C. M., Linn S. Purification and characterization of an apurinic/apyrimidinic endonuclease from HeLa cells. J Biol Chem. 1981 Apr 10;256(7):3405–3414. [PubMed] [Google Scholar]
- Katcher H. L., Wallace S. S. Characterization of the Escherichia coli X-ray endonuclease, endonuclease III. Biochemistry. 1983 Aug 16;22(17):4071–4081. doi: 10.1021/bi00286a013. [DOI] [PubMed] [Google Scholar]
- Kuhnlein U., Lee B., Linn S. Human uracil DNA N-glycosidase: studies in normal and repair defective cultured fibroblasts. Nucleic Acids Res. 1978 Jan;5(1):117–125. doi: 10.1093/nar/5.1.117. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kuhnlein U., Penhoet E. E., Linn S. An altered apurinic DNA endonuclease activity in group A and group D xeroderma pigmentosum fibroblasts. Proc Natl Acad Sci U S A. 1976 Apr;73(4):1169–1173. doi: 10.1073/pnas.73.4.1169. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Margison G. P., Pegg A. E. Enzymatic release of 7-methylguanine from methylated DNA by rodent liver extracts. Proc Natl Acad Sci U S A. 1981 Feb;78(2):861–865. doi: 10.1073/pnas.78.2.861. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nes I. F. Purification and properties of a mouse-cell DNA-repair endonuclease, which recognizes lesions in DNA induced by ultraviolet light, depurination, gamma-rays, and OsO4 treatment. Eur J Biochem. 1980 Nov;112(1):161–168. doi: 10.1111/j.1432-1033.1980.tb04997.x. [DOI] [PubMed] [Google Scholar]
- Pero J., Hannett N. M., Talkington C. Restriction cleavage map of SP01 DNA: general location of early, middle, and late genes. J Virol. 1979 Jul;31(1):156–171. doi: 10.1128/jvi.31.1.156-171.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sirover M. A. Induction of the DNA repair enzyme uracil-DNA glycosylase in stimulated human lymphocytes. Cancer Res. 1979 Jun;39(6 Pt 1):2090–2095. [PubMed] [Google Scholar]
- Strider W., Camien M. N., Warner R. C. Renaturation of denatured, covalently closed circular DNA. J Biol Chem. 1981 Aug 10;256(15):7820–7829. [PubMed] [Google Scholar]
- Teebor G. W., Frenkel K., Goldstein M. S. Ionizing radiation and tritium transmutation both cause formation of 5-hydroxymethyl-2'-deoxyuridine in cellular DNA. Proc Natl Acad Sci U S A. 1984 Jan;81(2):318–321. doi: 10.1073/pnas.81.2.318. [DOI] [PMC free article] [PubMed] [Google Scholar]
- van Lancker J. L., Tomura T. Purification and some properties of a mammalian repair endonuclease. Biochim Biophys Acta. 1974 Jun 14;353(1):99–114. doi: 10.1016/0005-2787(74)90101-4. [DOI] [PubMed] [Google Scholar]