Influence of minor groove binders on the eukaryotic topoisomerase II cleavage reaction with 41 base pair model oligonucleotides |
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Authors: | Achim Bell Leonhard Kittler Günter Löber Christoph Zimmer |
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Affiliation: | (1) Institut für Molekulare Biotechnologie e.V., Beutenbergstr. 11, D-07745 Jena, Germany;(2) Institut für Molekularbiologie, Friedrich-Schiller-Universität Jena, Winzerlaer Str. 10, D-07745 Jena, Germany |
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Abstract: | Summary This report deals with the cleavage reaction of calf thymus (CT) topoisomerase II with oligonucleotides containing one main cleavage site and adjacent binding sites for minor groove binders. The sequences of the oligonucleotides were derived from a pBR 322 sequence, which contains one main topoisomerase II cleavage site. The cleavage reaction was performed under increasing concentrations of minor groove binders and it showed characteristic inhibition dependences of topoisomerase II to the binding sites and to the binding length of the minor groove binders. The extension of the minor groove binder length on DNA from 4 to 10 base pairs (bp) by netropsin and bis-netropsin, respectively, causes a strong increase of the topoisomerase II cleavage inhibition. The same is observed by the introduction of a second minor groove binder sequence symmetrically positioned around the topoisomerase II main cleavage site. The combination of two different minor groove binders can lead to an increased topoisomerase II inhibition but also to a prevention of total inhibition as shown with chromomycin A3 and distamycin A at concentrations of 0.1 and 0.25 M, respectively. |
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Keywords: | model oligonucleotide minor groove binder CT topoisomerase II cleavage process |
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