Sequence-dependent effect of camptothecin on human topoisomerase I DNA cleavage

This article is dedicated to Arthur Kornberg on the occasion of his 70th birthday by a former post-doctoral fellow (O.W.) from his laboratory.
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Abstract

We have studied the effect of the antitumor drug, camptothecin, on the interaction of human topoisomerase I with DNA at the sequence level. At a low molar ratio of enzyme to DNA, cleavage is prominent and unique, located at a previously described hexadecameric recognition sequence, while a number of strong additional cleavage sites appear in the presence of the drug. Camptothecin stimulates cleavage at the recognition sequence less than twofold, whereas cleavage at the additional sites is stimulated up to 200-fold. Camptothecin greatly enhances the stability of the cleavable complexes formed at the additional sites, whereas the complex formed at the hexadecameric sequence is only marginally affected. Cleavage was eliminated at certain sites in the presence of camptothecin. Taken together these observations demonstrate that at least three types of potential eukaryotic topoisomerase I cleavage sites can be distinguished by the use of camptothecin. Comparison of the sequences at the additional cleavage sites in the presence of camptothecin reveals that the most frequently cleaved dinucleotide is TG with no consensus for the flanking nucleotides.

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    This study was supported by the Commission of the European Communities (contract B-6-0170-DK), the Danish Medical Research Council (grant no. 5.10.17.06), the Danish Cancer Society (grant nos 86-178, 87-025, and 87-088), the National Agency of Technology (contract no. 1985-133/001-85.521), and the Danish Programme of Biotechnology.

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