title: A repair competition assay to assess recognition by human nucleotide excision repair.
abstract: We developed a competition assay to compare, in a quantitative manner, the ability of human nucleotide excision repair (NER) to recognise structurally different forms of DNA damage. This assay uses a NER substrate consisting of M13 double-stranded DNA with a single and uniquely located acetylaminofluorene (AAF) adduct, and measures the efficiency by which multiply damaged plasmid DNA competes for excision repair of the site-directed modification. To validate this assay, we tested competitor DNA containing defined numbers of either AAF adducts or UV radiation products. In both cases, repair of the site-directed NER substrate was inhibited in a damage-specific and dose-dependent manner. We then exploited this competition assay to determine the susceptibility of bulky adozelesin-DNA adducts to human NER.
authors.affiliation: Institute of Pharmacology and Toxicology, University of Zürich-Tierspital, Zürich, Switzerland.
authors.forename: M T
authors.initials: MT
authors.lastname: Hess
authors.forename: D
authors.initials: D
authors.lastname: Gunz
authors.forename: H
authors.initials: H
authors.lastname: Naegeli
mesh_terms: D001483:Base Sequence; D001572:Benzofurans; D001667:Binding, Competitive; D003509:Cyclohexanecarboxylic Acids; D053138:Cyclohexenes; D004249:DNA Damage; D004260:DNA Repair; D000080890:Duocarmycins; D006801:Humans; D007211:Indoles; D015336:Molecular Probe Techniques; D008969:Molecular Sequence Data
chemical_list: D001572:Benzofurans; D003509:Cyclohexanecarboxylic Acids; D053138:Cyclohexenes; D000080890:Duocarmycins; D007211:Indoles; C060090:adozelesin