Anion-dependent modulations of DNA topoisomerase II-mediated reactions in potassium-containing solutions.
DNA binding proteins operate in an intracellular environment of low chloride concentration, yet in vitro assays of the activities of these proteins are often performed in isotonic chloride-containing solutions. Previously, the activity of bacterial DNA-binding proteins was found to be enhanced in potassium-containing solutions in which the anion glutamate (Glu) was substituted for chloride. We have extended this observation to include eukaryotic topoisomerase I and II activities. The concentration ranges over which DNA strand passing activities of these enzymes were observed was broader in KGlu than in KCl. This was also true for the topoisomerase II-mediated DNA strand passage and antineoplastic drug-dependent DNA cleavage produced by nuclear extracts from HL-60 human leukemia cells. The rate of topoisomerase II-mediated DNA strand passage was also dependent on anion moiety and concentration in potassium-containing buffers. Drug-dependent topoisomerase II-mediated DNA cleavage in intact HL-60 cell nuclei was also anion-dependent, suggesting that anion type and concentration may influence topoisomerase II-mediated events in mammalian cells as had been described for other DNA binding proteins in prokaryotic systems. This should be considered in developing biochemical assays of topoisomerase activities to reproduce intracellular conditions.
D000838:Anions; D002460:Cell Line; D002467:Cell Nucleus / Q000201:enzymology / Q000378:metabolism; D004249:DNA Damage; D004264:DNA Topoisomerases, Type I / Q000378:metabolism / Q000502:physiology; D004250:DNA Topoisomerases, Type II / Q000378:metabolism* / Q000502:physiology; D005971:Glutamates; D018698:Glutamic Acid; D006801:Humans; D007942:Leukemia, Experimental / Q000201:enzymology; D011189:Potassium Chloride; D012996:Solutions
D000838:Anions; D005971:Glutamates; D012996:Solutions; D018698:Glutamic Acid; D011189:Potassium Chloride; D004264:DNA Topoisomerases, Type I; D004250:DNA Topoisomerases, Type II