title: 7,8-Dihydro-8-oxoadenine, a highly mutagenic adduct, is repaired by Escherichia coli and human mismatch-specific uracil/thymine-DNA glycosylases.
abstract: Hydroxyl radicals predominantly react with the C(8) of purines forming 7,8-dihydro-8-oxoguanine (8oxoG) and 7,8-dihydro-8-oxoadenine (8oxoA) adducts, which are highly mutagenic in mammalian cells. The majority of oxidized DNA bases are removed by DNA glycosylases in the base excision repair pathway. Here, we report for the first time that human thymine-DNA glycosylase (hTDG) and Escherichia coli mismatch-specific uracil-DNA glycosylase (MUG) can remove 8oxoA from 8oxoA•T, 8oxoA•G and 8oxoA•C pairs. Comparison of the kinetic parameters of the reaction indicates that full-length hTDG excises 8oxoA, 3,N(4)-ethenocytosine (εC) and T with similar efficiency (k(max) = 0.35, 0.36 and 0.16 min(-1), respectively) and is more proficient as compared with its bacterial homologue MUG. The N-terminal domain of the hTDG protein is essential for 8oxoA-DNA glycosylase activity, but not for εC repair. Interestingly, the TDG status had little or no effect on the proliferation rate of mouse embryonic fibroblasts after exposure to γ-irradiation. Nevertheless, using whole cell-free extracts from the DNA glycosylase-deficient murine embryonic fibroblasts and E. coli, we demonstrate that the excision of 8oxoA from 8oxoA•T and 8oxoA•G has an absolute requirement for TDG and MUG, respectively. The data establish that MUG and TDG can counteract the genotoxic effects of 8oxoA residues in vivo.
authors.affiliation: Groupe Réparation de l'ADN, Université Paris Sud, Laboratoire Stabilité Génétique et Oncogenèse CNRS, UMR 8200, Institut de Cancérologie Gustave Roussy, F-94805 Villejuif Cedex, France.
authors.forename: Ibtissam
authors.initials: I
authors.lastname: Talhaoui
authors.forename: Sophie
authors.initials: S
authors.lastname: Couvé
authors.forename: Alexander A
authors.initials: AA
authors.lastname: Ishchenko
authors.forename: Christophe
authors.initials: C
authors.lastname: Kunz
authors.forename: Primo
authors.initials: P
authors.lastname: Schär
authors.forename: Murat
authors.initials: M
authors.lastname: Saparbaev
mesh_terms: D000225:Adenine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D020029:Base Pairing; D002460:Cell Line; D018736:DNA Adducts / Q000737:chemistry / Q000378:metabolism*; D004260:DNA Repair; D004926:Escherichia coli / Q000201:enzymology; D006801:Humans; D051379:Mice; D016296:Mutagenesis; D011839:Radiation, Ionizing; D013941:Thymine / Q000737:chemistry; D044502:Thymine DNA Glycosylase / Q000378:metabolism*
chemical_list: D018736:DNA Adducts; C002992:8-hydroxyadenine; D044502:Thymine DNA Glycosylase; C113921:mismatch-specific thymine uracil-DNA glycosylase; D000225:Adenine; D013941:Thymine