The H circles of Leishmania tarentolae are a unique amplifiable system of oligomeric DNAs associated with drug resistance.
We have induced drug resistance against methotrexate, an inhibitor of dihydrofolate reductase, and CB3717, an inhibitor of thymidylate synthetase in a strain of Leishmania tarentolae. The drug-resistant strains contain extrachromosomal DNA circles of 68 kilobases with a 30-kilobase inverted duplication flanked by 4- and 5 kilobase unique segments. We show that these circles are highly homologous to the drug-induced H circles of L. tropica (1). All three L. tarentolae strains analyzed contain a chromosomal copy of the H region without duplication, but two of the three strains contain extrachromosomal H circles as well, predominantly present as H circle dimers in one strain and as tetramers in the other. After induction of methotrexate resistance, monomeric circles, presumably derived from the oligomers, become the major type of circle. Our results indicate that the H region represents a genomic region that can be copied at very low frequency to yield circles by a precise, but unusual mechanism under natural conditions in wild-type cells. Although superficially analogous to the episomes of bacteria, the system is without precedent in nature.
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Division of Molecular Biology, Netherlands Cancer Institute, Antoni van Leeuwenhoekhuis, Amsterdam.
Fase-Fowler
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van Luenen
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Calafat
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Borst
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D000818:Animals; D003001:Cloning, Molecular; D004247:DNA / Q000032:analysis* / Q000648:ultrastructure; D004270:DNA, Circular / Q000187:drug effects*; D004351:Drug Resistance; D005492:Folic Acid / Q000031:analogs & derivatives* / Q000494:pharmacology; D007891:Leishmania / Q000235:genetics*; D008727:Methotrexate / Q000494:pharmacology*; D008854:Microscopy, Electron; D011799:Quinazolines / Q000494:pharmacology*; D013940:Thymidylate Synthase / Q000037:antagonists & inhibitors
D004270:DNA, Circular; D011799:Quinazolines; C031662:CB 3717; D004247:DNA; D005492:Folic Acid; D013940:Thymidylate Synthase; D008727:Methotrexate