DNA topoisomerases from pathogenic fungi: targets for the discovery of antifungal drugs.
DNA topoisomerases, a class of enzymes that change the topological structure of DNA, have been shown to be the target of many therapeutic agents, including antibacterial agents (quinolones) and anticancer agents. These drugs inhibit the enzyme in a unique way so that the enzyme is converted into a cellular poison. Candida albicans and Aspergillus niger are two major opportunistic fungal pathogens. Our results show that these fungi have high levels of both type I and type II topoisomerases (with a minimum of 5 x 10(5) ATP-independent relaxation units and 2 x 10(5) P-4 unknotting units per liter of wild-type C. albicans). The ATP-dependent type II topoisomerase (termed C. albicans topoisomerase II) was purified by approximately 2,000-fold from C. albicans cells by using a simple isolation scheme that consists of three column procedures: hydroxylapatite, phosphocellulose, and heparin-agarose chromatographies. The responses of the Candida and the calf thymus topoisomerase II to some known topoisomerase II inhibitors were measured. Etoposide and 4'-(9-acridinylamino)methanesulfon-m-anisidide, compounds known to inhibit catalysis and to enhance DNA breakage by mammalian topoisomerase II, and A-80198, an etoposide derivative, enhanced cleavage by both enzymes at similar concentrations of these compounds, with the response of the calf thymus topoisomerase II from slightly to fourfold higher in magnitude than the response of the Candida enzyme in the same concentration range. In contrast, A-75272 (a cytotoxic tricyclic quinolone) shows a slightly stronger DNA cleavage enhancement effect with the Candida enzyme than with the mammalian counterpart. The abundance of the enzyme in cells and the different drug responses of the host enzyme and the fungal enzyme suggest that the fungal topoisomerase may serve as a target for the discovery of effective and safe antifungal agents.
Shen
L L
LL
Anti-Infective Research Division, Abbott Laboratories, Abbott Park, Illinois 60064, USA.
Baranowski
J
J
Fostel
J
J
Montgomery
D A
DA
Lartey
P A
PA
D000818:Animals; D000935:Antifungal Agents / Q000494:pharmacology*; D001234:Aspergillus niger / Q000187:drug effects* / Q000201:enzymology*; D002176:Candida albicans / Q000187:drug effects* / Q000201:enzymology*; D002417:Cattle; D004264:DNA Topoisomerases, Type I / Q000187:drug effects* / Q000302:isolation & purification; D004250:DNA Topoisomerases, Type II / Q000187:drug effects* / Q000302:isolation & purification; D009181:Mycoses / Q000188:drug therapy / Q000382:microbiology; D059004:Topoisomerase I Inhibitors; D059005:Topoisomerase II Inhibitors
D000935:Antifungal Agents; D059004:Topoisomerase I Inhibitors; D059005:Topoisomerase II Inhibitors; D004264:DNA Topoisomerases, Type I; D004250:DNA Topoisomerases, Type II