Effects of uracil incorporation, DNA mismatches, and abasic sites on cleavage and religation activities of mammalian topoisomerase I.
Abasic sites and deamination of cytosine to uracil are probably the most common types of endogenous DNA damage. The effects of such lesions on DNA topoisomerase I (top1) activity were examined in oligonucleotides containing a unique top1 cleavage site. The presence of uracils and abasic sites within the first 4 bases immediately 5' to the cleavage site suppressed normal top1 cleavage and induced new top1 cleavage sites. Uracils immediately 3' to the cleavage site increased cleavage and produced a camptothecin mimicking effect. A mismatch with a bulge or abasic sites immediately 3' to the top1 cleavage site irreversibly trapped top1 cleavable complexes in the absence of camptothecin and produced a suicide cleavage complex. These results demonstrate that top1 activity is sensitive to physiological, environmental, and pharmacological DNA modifications and that top1 can act as a specific mismatch- and abasic site-nicking enzyme.
Pourquier
P
P
Laboratory of Molecular Pharmacology, Division of Basic Sciences, NCI, National Institutes of Health, Bethesda, Maryland 20892, USA.
Ueng
L M
LM
Kohlhagen
G
G
Mazumder
A
A
Gupta
M
M
Kohn
K W
KW
Pommier
Y
Y
D000818:Animals; D001665:Binding Sites; D004247:DNA / Q000235:genetics*; D004264:DNA Topoisomerases, Type I / Q000378:metabolism*; D006868:Hydrolysis; D008322:Mammals; D009692:Nucleic Acid Heteroduplexes; D014498:Uracil / Q000378:metabolism*
D009692:Nucleic Acid Heteroduplexes; D014498:Uracil; D004247:DNA; D004264:DNA Topoisomerases, Type I