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Nucleic Acids Research, Vol 24, Issue 5 824-828, Copyright © 1996 by Oxford University Press

ARTICLES

A repair competition assay to assess recognition by human nucleotide excision repair

MT Hess, D Gunz and H Naegeli
Institute of Pharmacology and Toxicology, University of Zurich- Tierspital, Zurich, Switzerland.

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.
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