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. 1994 Sep 11;22(18):3760–3764. doi: 10.1093/nar/22.18.3760

Formamidopyrimidine DNA glycosylase in the yeast Saccharomyces cerevisiae.

R de Oliveira 1, P A van der Kemp 1, D Thomas 1, A Geiger 1, P Nehls 1, S Boiteux 1
PMCID: PMC308359  PMID: 7937089

Abstract

A DNA glycosylase that excises, 2,6-diamino-4-hydroxy-5N-methylformamidopyrimidine (Fapy) from double stranded DNA has been purified 28,570-fold from the yeast Saccharomyces cerevisiae. Gel filtration chromatography shows that yeast Fapy DNA glycosylase has a molecular weight of about 40 kDa. The Fapy DNA glycosylase is active in the presence of EDTA, but is completely inhibited by 0.2 M KCl. Yeast Fapy DNA glycosylase does not excise N7-methylguanine, N3-methyladenine or uracil. A repair enzyme for 7,8-dihydro-8-oxoguanine (8-OxoG) co-purifies with the Fapy DNA glycosylase. This repair activity causes strand cleavage at the site of 8-OxoG in DNA duplexes. The highest rate of incision of the 8-OxoG-containing strand was observed for duplexes where 8-OxoG was opposite guanine. The mode of incision at 8-OxoG was not established yet. The results however suggest that the Fapy- and 8-OxoG-repair activities are associated with a single protein.

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Selected References

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

  1. Ames B. N., Shigenaga M. K., Hagen T. M. Oxidants, antioxidants, and the degenerative diseases of aging. Proc Natl Acad Sci U S A. 1993 Sep 1;90(17):7915–7922. doi: 10.1073/pnas.90.17.7915. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bailly V., Verly W. G., O'Connor T., Laval J. Mechanism of DNA strand nicking at apurinic/apyrimidinic sites by Escherichia coli [formamidopyrimidine]DNA glycosylase. Biochem J. 1989 Sep 1;262(2):581–589. doi: 10.1042/bj2620581. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Berdal K. G., Bjørås M., Bjelland S., Seeberg E. Cloning and expression in Escherichia coli of a gene for an alkylbase DNA glycosylase from Saccharomyces cerevisiae; a homologue to the bacterial alkA gene. EMBO J. 1990 Dec;9(13):4563–4568. doi: 10.1002/j.1460-2075.1990.tb07909.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bessho T., Tano K., Kasai H., Ohtsuka E., Nishimura S. Evidence for two DNA repair enzymes for 8-hydroxyguanine (7,8-dihydro-8-oxoguanine) in human cells. J Biol Chem. 1993 Sep 15;268(26):19416–19421. [PubMed] [Google Scholar]
  5. Boiteux S., Belleney J., Roques B. P., Laval J. Two rotameric forms of open ring 7-methylguanine are present in alkylated polynucleotides. Nucleic Acids Res. 1984 Jul 11;12(13):5429–5439. doi: 10.1093/nar/12.13.5429. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Boiteux S., Gajewski E., Laval J., Dizdaroglu M. Substrate specificity of the Escherichia coli Fpg protein (formamidopyrimidine-DNA glycosylase): excision of purine lesions in DNA produced by ionizing radiation or photosensitization. Biochemistry. 1992 Jan 14;31(1):106–110. doi: 10.1021/bi00116a016. [DOI] [PubMed] [Google Scholar]
  7. Boiteux S., Huisman O. Isolation of a formamidopyrimidine-DNA glycosylase (fpg) mutant of Escherichia coli K12. Mol Gen Genet. 1989 Jan;215(2):300–305. doi: 10.1007/BF00339732. [DOI] [PubMed] [Google Scholar]
  8. Boiteux S., O'Connor T. R., Laval J. Formamidopyrimidine-DNA glycosylase of Escherichia coli: cloning and sequencing of the fpg structural gene and overproduction of the protein. EMBO J. 1987 Oct;6(10):3177–3183. doi: 10.1002/j.1460-2075.1987.tb02629.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Boiteux S., O'Connor T. R., Lederer F., Gouyette A., Laval J. Homogeneous Escherichia coli FPG protein. A DNA glycosylase which excises imidazole ring-opened purines and nicks DNA at apurinic/apyrimidinic sites. J Biol Chem. 1990 Mar 5;265(7):3916–3922. [PubMed] [Google Scholar]
  10. Boiteux S. Properties and biological functions of the NTH and FPG proteins of Escherichia coli: two DNA glycosylases that repair oxidative damage in DNA. J Photochem Photobiol B. 1993 Jul;19(2):87–96. doi: 10.1016/1011-1344(93)87101-r. [DOI] [PubMed] [Google Scholar]
  11. Breimer L. H. Enzymatic excision from gamma-irradiated polydeoxyribonucleotides of adenine residues whose imidazole rings have been ruptured. Nucleic Acids Res. 1984 Aug 24;12(16):6359–6367. doi: 10.1093/nar/12.16.6359. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Castaing B., Geiger A., Seliger H., Nehls P., Laval J., Zelwer C., Boiteux S. Cleavage and binding of a DNA fragment containing a single 8-oxoguanine by wild type and mutant FPG proteins. Nucleic Acids Res. 1993 Jun 25;21(12):2899–2905. doi: 10.1093/nar/21.12.2899. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Chen J., Derfler B., Maskati A., Samson L. Cloning a eukaryotic DNA glycosylase repair gene by the suppression of a DNA repair defect in Escherichia coli. Proc Natl Acad Sci U S A. 1989 Oct;86(20):7961–7965. doi: 10.1073/pnas.86.20.7961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. Crosby B., Prakash L., Davis H., Hinkle D. C. Purification and characterization of a uracil-DNA glycosylase from the yeast. Saccharomyces cerevisiae. Nucleic Acids Res. 1981 Nov 11;9(21):5797–5809. doi: 10.1093/nar/9.21.5797. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Czeczot H., Tudek B., Lambert B., Laval J., Boiteux S. Escherichia coli Fpg protein and UvrABC endonuclease repair DNA damage induced by methylene blue plus visible light in vivo and in vitro. J Bacteriol. 1991 Jun;173(11):3419–3424. doi: 10.1128/jb.173.11.3419-3424.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Dianov G., Price A., Lindahl T. Generation of single-nucleotide repair patches following excision of uracil residues from DNA. Mol Cell Biol. 1992 Apr;12(4):1605–1612. doi: 10.1128/mcb.12.4.1605. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Gossett J., Lee K., Cunningham R. P., Doetsch P. W. Yeast redoxyendonuclease, a DNA repair enzyme similar to Escherichia coli endonuclease III. Biochemistry. 1988 Apr 5;27(7):2629–2634. doi: 10.1021/bi00407a054. [DOI] [PubMed] [Google Scholar]
  20. Grollman A. P., Moriya M. Mutagenesis by 8-oxoguanine: an enemy within. Trends Genet. 1993 Jul;9(7):246–249. doi: 10.1016/0168-9525(93)90089-z. [DOI] [PubMed] [Google Scholar]
  21. Hamilton K. K., Kim P. M., Doetsch P. W. A eukaryotic DNA glycosylase/lyase recognizing ultraviolet light-induced pyrimidine dimers. Nature. 1992 Apr 23;356(6371):725–728. doi: 10.1038/356725a0. [DOI] [PubMed] [Google Scholar]
  22. Hoeijmakers J. H. Nucleotide excision repair. II: From yeast to mammals. Trends Genet. 1993 Jun;9(6):211–217. doi: 10.1016/0168-9525(93)90121-w. [DOI] [PubMed] [Google Scholar]
  23. Lindahl T. Instability and decay of the primary structure of DNA. Nature. 1993 Apr 22;362(6422):709–715. doi: 10.1038/362709a0. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. Michaels M. L., Cruz C., Grollman A. P., Miller J. H. Evidence that MutY and MutM combine to prevent mutations by an oxidatively damaged form of guanine in DNA. Proc Natl Acad Sci U S A. 1992 Aug 1;89(15):7022–7025. doi: 10.1073/pnas.89.15.7022. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. O'Connor T. R., Laval J. Human cDNA expressing a functional DNA glycosylase excising 3-methyladenine and 7-methylguanine. Biochem Biophys Res Commun. 1991 May 15;176(3):1170–1177. doi: 10.1016/0006-291x(91)90408-y. [DOI] [PubMed] [Google Scholar]
  27. Olsen L. C., Aasland R., Wittwer C. U., Krokan H. E., Helland D. E. Molecular cloning of human uracil-DNA glycosylase, a highly conserved DNA repair enzyme. EMBO J. 1989 Oct;8(10):3121–3125. doi: 10.1002/j.1460-2075.1989.tb08464.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Ortigão J. F., Rösch H., Selter H., Fröhlich A., Lorenz A., Montenarh M., Seliger H. Antisense effect of oligodeoxynucleotides with inverted terminal internucleotidic linkages: a minimal modification protecting against nucleolytic degradation. Antisense Res Dev. 1992 Summer;2(2):129–146. doi: 10.1089/ard.1992.2.129. [DOI] [PubMed] [Google Scholar]
  29. Percival K. J., Klein M. B., Burgers P. M. Molecular cloning and primary structure of the uracil-DNA-glycosylase gene from Saccharomyces cerevisiae. J Biol Chem. 1989 Feb 15;264(5):2593–2598. [PubMed] [Google Scholar]
  30. Sancar A., Sancar G. B. DNA repair enzymes. Annu Rev Biochem. 1988;57:29–67. doi: 10.1146/annurev.bi.57.070188.000333. [DOI] [PubMed] [Google Scholar]
  31. Sancar A., Tang M. S. Nucleotide excision repair. Photochem Photobiol. 1993 May;57(5):905–921. doi: 10.1111/j.1751-1097.1993.tb09233.x. [DOI] [PubMed] [Google Scholar]
  32. Tchou J., Kasai H., Shibutani S., Chung M. H., Laval J., Grollman A. P., Nishimura S. 8-oxoguanine (8-hydroxyguanine) DNA glycosylase and its substrate specificity. Proc Natl Acad Sci U S A. 1991 Jun 1;88(11):4690–4694. doi: 10.1073/pnas.88.11.4690. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Tudek B., Laval J., Boiteux S. SOS-independent mutagenesis in lacZ induced by methylene blue plus visible light. Mol Gen Genet. 1993 Jan;236(2-3):433–439. doi: 10.1007/BF00277144. [DOI] [PubMed] [Google Scholar]

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