Selective cleavage of kinetoplast DNA minicircles promoted by antitrypanosomal drugs.
Pentamidine, diminazene aceturate (Berenil), isometamidium chloride (Samorin), and ethidium bromide, which are important antitrypanosomal drugs, promote linearization of Trypanosoma equiperdum minicircle DNA (the principal component of kinetoplast DNA, the mitochondrial DNA in these parasites). This effect occurs at therapeutically relevant concentrations. The linearized minicircles are protease sensitive and are not digested by lambda exonuclease (a 5' to 3' exonuclease), indicating that the break is double stranded and that protein is bound to both 5' ends of the molecule. The cleavage sites map to discrete positions in the minicircle sequence, and the cleavage pattern varies with different drugs. These findings are characteristic for type II topoisomerase inhibitors, and they mimic the effects of the antitumor drug etoposide (VP16-213, a semisynthetic podophyllotoxin analog) on T. equiperdum minicircles. However, the antitrypanosomal drugs differ dramatically from etoposide in that they do not promote detectable formation of nuclear DNA-protein complexes or of strand breaks in nuclear DNA. Selective inhibition of a mitochondrial type II topoisomerase may explain why these antitrypanosomal drugs preferentially disrupt mitochondrial DNA structure and generate dyskinetoplastic trypanosomes (which lack mitochondrial DNA).
Shapiro
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Department of Biological Chemistry, Johns Hopkins University School of Medicine, Baltimore, MD 21205.
Englund
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D000818:Animals; D004264:DNA Topoisomerases, Type I / Q000378:metabolism; D004270:DNA, Circular / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D018105:DNA, Kinetoplast; D004133:Diminazene / Q000031:analogs & derivatives / Q000494:pharmacology; D004996:Ethidium / Q000494:pharmacology; D015394:Molecular Structure; D010419:Pentamidine / Q000494:pharmacology; D010617:Phenanthridines / Q000494:pharmacology; D015183:Restriction Mapping; D014344:Trypanocidal Agents / Q000494:pharmacology*; D014345:Trypanosoma / Q000187:drug effects / Q000235:genetics
D004270:DNA, Circular; D018105:DNA, Kinetoplast; D010617:Phenanthridines; D014344:Trypanocidal Agents; D010419:Pentamidine; C000702:isometamidium chloride; D004264:DNA Topoisomerases, Type I; D004996:Ethidium; C003915:diminazene aceturate; D004133:Diminazene