Response of Purified Mitochondrial DNA Topoisomerase I from Bovine Liver to Camptothecin and m-AMSA

Abstract

The type I DNA topoisomerase isolated from bovine liver mitochondria is demonstrated here to be inhibited by camptothecin, a plant alkaloid previously shown to target the nuclear type I topoisomerase in mammalian cells. The antitumor drug reduces the ability of the mitochondrial enzyme to relax positive as well as negative supercoils although the inhibition of the former process requires more than 60-fold more drug than the latter process. A similar response is seen with the nuclear topoisomerase I. Camptothecin also stimulates the mitochondrial topoisomerase-induced cleavage of pUC19 at numerous, discrete sites. The antitumor drug 4′-(9-acridinylamino)-methanesulfon-m-anisidide, which has been shown to target the nuclear topoisomerase II, inhibited the mitochondrial type I topoisomerase relaxation activity, but this effect was found to be the result of the drug intercalating into the negatively supercoiled DNA rather than from a specific interaction with the mitochondrial enzyme. VM-26, a nonintercalating topoisomerase II poison, showed no inhibitory effect up to a concentration of 50 μM.