Mammalian topoisomerase II activity is modulated by the DNA minor groove binder distamycin in simian virus 40 DNA.
DNA topoisomerases II are nuclear enzymes that have been identified recently as targets for some of the most active anticancer drugs. Antitumor topoisomerase II inhibitors such as teniposide (VM-26) produce enzyme-induced DNA cleavage and inhibition of enzyme activity. By adding to such reactions distamycin, a compound whose effects on DNA have been extensively characterized, we investigated the effects of drug binding upon topoisomerase II-mediated DNA cleavage induced by VM-26. We have found a correspondence between distamycin binding (determined by footprinting analysis) and topoisomerase II-mediated cleavage of SV40 DNA (determined by sequencing gel analysis). Distamycin binding potentiated the cleavage of specific sites in the near proximity of distamycin-binding sites (within at least 25 base pairs), which indicates that DNA secondary structure is involved in topoisomerase II-DNA interactions. That distamycin potentiated cleavage only at sites that were recognized in the absence of distamycin and suppressed cleavage directly at distamycin-binding sites indicates that topoisomerase II recognizes DNA on the basis of primary sequence. In addition, distamycin stimulated topoisomerase II-mediated DNA relaxation and antagonized the inhibitory effect of VM-26. These results show that the DNA sequence-specific binding of distamycin produces local and propagated effects in the DNA which markedly affect topoisomerase II activity.
Fesen
M
M
Laboratory of Molecular Pharmacology, National Cancer Institute, Bethesda, Maryland 20892.
Pommier
Y
Y
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D004262:DNA Restriction Enzymes; D004250:DNA Topoisomerases, Type II / Q000378:metabolism*; D004279:DNA, Viral / Q000378:metabolism*; D004214:Distamycins / Q000378:metabolism / Q000494:pharmacology*; D004347:Drug Interactions; D004586:Electrophoresis; D007939:Leukemia L1210 / Q000201:enzymology; D051379:Mice; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation / Q000187:drug effects; D011758:Pyrroles / Q000494:pharmacology*; D012091:Repetitive Sequences, Nucleic Acid; D013539:Simian virus 40 / Q000235:genetics*; D013713:Teniposide / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D004279:DNA, Viral; D004214:Distamycins; D011758:Pyrroles; D013713:Teniposide; D004262:DNA Restriction Enzymes; D004250:DNA Topoisomerases, Type II