Substrate specificity of the Escherichia coli Fpg protein (formamidopyrimidine-DNA glycosylase): excision of purine lesions in DNA produced by ionizing radiation or photosensitization
- PMID: 1731864
- DOI: 10.1021/bi00116a016
Substrate specificity of the Escherichia coli Fpg protein (formamidopyrimidine-DNA glycosylase): excision of purine lesions in DNA produced by ionizing radiation or photosensitization
Abstract
We have investigated the excision of a variety of modified bases from DNA by the Escherichia coli Fpg protein (formamidopyrimidine-DNA glycosylase) [Boiteux, S., O'Connor, T. R., Lederer, F., Gouyette, A., & Laval, J. (1990) J. Biol. Chem. 265, 3916-3922]. DNA used as a substrate was modified either by exposure to ionizing radiation or by photosensitization using visible light in the presence of methylene blue (MB). The technique of gas chromatography/mass spectrometry, which can unambiguously identify and quantitate pyrimidine- and purine-derived lesions in DNA, was used for analysis of hydrolyzed and derivatized DNA samples. Thirteen products resulting from pyrimidines and purines were detected in gamma-irradiated DNA, whereas only the formation of 2,6-diamino-4-hydroxy-5-formamidopyrimidine (FapyGua) and 8-hydroxyguanine (8-OH-Gua) was observed in visible light/MB-treated DNA. Analysis of gamma-irradiated DNA after incubation with the Fpg protein followed by precipitation revealed that the Fpg protein significantly excised 4,6-diamino-5-formamidopyrimidine (FapyAde), FapyGua, and 8-OH-Gua. The excision of a small but detectable amount of 8-hydroxyadenine was also observed. The detection of these products in the supernatant fractions of the same samples confirmed their excision by the enzyme. Nine pyrimidine-derived lesions were not excised. The Fpg protein also excised FapyGua and 8-OH-Gua from visible light/MB-treated DNA. The presence of these products in the supernatant fractions confirmed their excision.(ABSTRACT TRUNCATED AT 250 WORDS)
Similar articles
-
Substrate specificity of Deinococcus radiodurans Fpg protein.Biochemistry. 1999 Jul 20;38(29):9435-9. doi: 10.1021/bi990680m. Biochemistry. 1999. PMID: 10413519
-
Kinetics of excision of purine lesions from DNA by Escherichia coli Fpg protein.Nucleic Acids Res. 1997 Feb 1;25(3):474-9. doi: 10.1093/nar/25.3.474. Nucleic Acids Res. 1997. PMID: 9016584 Free PMC article.
-
Excision of oxidatively damaged DNA bases by the human alpha-hOgg1 protein and the polymorphic alpha-hOgg1(Ser326Cys) protein which is frequently found in human populations.Nucleic Acids Res. 1999 Oct 15;27(20):4001-7. doi: 10.1093/nar/27.20.4001. Nucleic Acids Res. 1999. PMID: 10497264 Free PMC article.
-
DNA substrates containing defined oxidative base lesions and their application to study substrate specificities of base excision repair enzymes.Prog Nucleic Acid Res Mol Biol. 2001;68:207-21. doi: 10.1016/s0079-6603(01)68101-7. Prog Nucleic Acid Res Mol Biol. 2001. PMID: 11554298 Review.
-
Imidazole ring-opened DNA purines and their biological significance.J Biochem Mol Biol. 2003 Jan 31;36(1):12-9. doi: 10.5483/bmbrep.2003.36.1.012. J Biochem Mol Biol. 2003. PMID: 12542970 Review.
Cited by
-
An OGG1 orthologue encoding a functional 8-oxoguanine DNA glycosylase/lyase in Arabidopsis thaliana.Plant Mol Biol. 2001 Dec;47(6):795-804. doi: 10.1023/a:1013644026132. Plant Mol Biol. 2001. PMID: 11785940
-
Repair of DNA lesions induced by hydrogen peroxide in the presence of iron chelators in Escherichia coli: participation of endonuclease IV and Fpg.J Bacteriol. 2000 Apr;182(7):1964-8. doi: 10.1128/JB.182.7.1964-1968.2000. J Bacteriol. 2000. PMID: 10715004 Free PMC article.
-
A genome-wide distribution of 8-oxoguanine correlates with the preferred regions for recombination and single nucleotide polymorphism in the human genome.Genome Res. 2006 May;16(5):567-75. doi: 10.1101/gr.4769606. Genome Res. 2006. PMID: 16651663 Free PMC article.
-
Genetic effects of oxidative DNA damages: comparative mutagenesis of the imidazole ring-opened formamidopyrimidines (Fapy lesions) and 8-oxo-purines in simian kidney cells.Nucleic Acids Res. 2006 May 5;34(8):2305-15. doi: 10.1093/nar/gkl099. Print 2006. Nucleic Acids Res. 2006. PMID: 16679449 Free PMC article.
-
Oxidative damage to DNA: do we have a reliable biomarker?Environ Health Perspect. 1996 May;104 Suppl 3(Suppl 3):465-9. doi: 10.1289/ehp.96104s3465. Environ Health Perspect. 1996. PMID: 8781365 Free PMC article.
Publication types
MeSH terms
Substances
LinkOut - more resources
Other Literature Sources
Molecular Biology Databases
Miscellaneous