Sensitivity of human type II topoisomerases to DNA damage: stimulation of enzyme-mediated DNA cleavage by abasic, oxidized and alkylated lesions.
Type II topoisomerases are essential enzymes that are also the primary cellular targets for a number of important anticancer drugs. These drugs act by increasing levels of topoisomerase II-mediated DNA cleavage. Recent studies indicate that endogenous forms of DNA damage, such as abasic sites and base mismatches, also stimulate the DNA scission activity of the enzyme. To extend our understanding of how type II topoisomerases react to DNA damage, the effects of abasic sites, and oxidized and alkylated bases on DNA cleavage mediated by human topo-isomerase IIalpha and beta were determined. Based on experiments that incorporated random abasic sites into plasmid DNA, human type II enzymes can locate lesions even within a background of several thousand undamaged base pairs. As determined by experiments that utilized site-specific forms of DNA lesions, oxidized or monoalkylated purines that allow base pairing and induce little distortion in the double helix have modest effects on topoisomerase II-mediated DNA cleavage. In contrast, 1,N(6)-ethenoadenine, a bulky lesion that disrupts base pairing, enhanced DNA cleavage approximately 10-fold. 1,N(6)-Ethenoadenine is the first lesion found to rival the stimulatory effects of apurinic sites on the DNA scission activity of eukaryotic type II topoisomerases.
Sabourin
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M
Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, TN 37232-0146, USA.
Osheroff
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N
D000225:Adenine / Q000031:analogs & derivatives / Q000378:metabolism; D000478:Alkylation; D001080:Apurinic Acid / Q000378:metabolism; D001483:Base Sequence; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D018736:DNA Adducts / Q000737:chemistry / Q000378:metabolism; D004249:DNA Damage; D004260:DNA Repair; D004250:DNA Topoisomerases, Type II / Q000378:metabolism*; D004268:DNA-Binding Proteins; D006147:Guanine / Q000031:analogs & derivatives / Q000378:metabolism; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism; D009154:Mutation; D009841:Oligonucleotides / Q000737:chemistry / Q000378:metabolism; D010084:Oxidation-Reduction; D010957:Plasmids / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012680:Sensitivity and Specificity
D000225:Adenine; D001080:Apurinic Acid; D004247:DNA; D018736:DNA Adducts; D004250:DNA Topoisomerases, Type II; D004268:DNA-Binding Proteins; D006147:Guanine; D007527:Isoenzymes; D009841:Oligonucleotides; C002992:8-hydroxyadenine; C024829:8-hydroxyguanine; C008449:O-(6)-methylguanine; C000730567:6-methyladenine