title: Sensitive analysis of oxidative DNA damage in mammalian cells: use of the bacterial Fpg protein in combination with alkaline unwinding.
abstract: The measurement of oxidative DNA base modifications by different methods has received special attention in recent years. Here we describe a procedure to quantify DNA lesions recognized by the bacterial formamido-pyrimidine-DNA glycosylases (Fpg protein). These include 7,8-dihydro-8-oxoguanine (8-hydroxyguanine) as well as some other forms of imidazole ring-opened purines, which are converted into abasic sites and subsequently into DNA single-strand breaks by the associated endonuclease activity. The frequency of DNA strand breaks is determined by the alkaline unwinding technique. The procedure provides a fast and sensitive tool to assess the extent of spontaneous as well as induced oxidative DNA damage in mammalian cells.
authors.affiliation: University of Bremen, Department of Biology and Chemistry, Germany.
authors.forename: A
authors.initials: A
authors.lastname: Hartwig
authors.forename: H
authors.initials: H
authors.lastname: Dally
authors.forename: R
authors.initials: R
authors.lastname: Schlepegrell
mesh_terms: D000818:Animals; D006224:Cricetinae; D003412:Cricetulus; D004247:DNA / Q000302:isolation & purification; D004249:DNA Damage / Q000187:drug effects / Q000502:physiology*; D004260:DNA Repair; D004277:DNA, Single-Stranded / Q000032:analysis* / Q000187:drug effects / Q000528:radiation effects; D043382:DNA-Formamidopyrimidine Glycosylase; D005347:Fibroblasts / Q000378:metabolism; D006367:HeLa Cells / Q000378:metabolism; D006801:Humans; D009699:N-Glycosyl Hydrolases / Q000378:metabolism*; D018384:Oxidative Stress / Q000502:physiology*
chemical_list: D004277:DNA, Single-Stranded; D004247:DNA; D009699:N-Glycosyl Hydrolases; D043382:DNA-Formamidopyrimidine Glycosylase