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A rapid and quantitative microtiter assay for eukaryotic topoisomerase II.

Topoisomerase II is required for decatenation of daughter chromatids during mitosis and has been shown to be an intracellular target by which certain antineoplastic drugs selectively kill tumor cells (for review, see 1,2). Identification of novel inhibitors requires large scale screening; however, most topo II assays (decatenation, DNA cleavage, etc.) are not ideal when extremely large numbers of samples must be evaluated. Here we describe a rapid and quantitative microtiter assay for topo II which is suitable for large scale screening of agents that alter topo II/DNA interaction. The assay is based upon the observation that catenated kinetoplast DNA (kDNA) is too large to diffuse out of an agarose plug whereas the decatenated topo II products freely diffuse out of the gel plug.