Stable amplified DNA in drug-resistant Leishmania exists as extrachromosomal circles.
The relative stability of amplified DNA in drug-resistant Leishmania major was previously reported to be dependent on location, that is, unstable amplified DNA was extrachromosomal and stable amplified DNA was chromosomal. Leishmanial chromosomes have now been directly examined by means of orthogonal-field-alternation gel electrophoresis (OFAGE). The amplified DNA's in three resistant cell lines displayed unusual migration and were clearly extrachromosomal, regardless of whether the amplified DNA's were stable or unstable. Thus, contrary to conclusions from earlier studies of drug resistance in cultured animal cells, stable amplified DNA in Leishmania can be extrachromosomal. In addition, these amplified DNA's were shown to be circular on the basis of their resistance to exonuclease III digestion and their behavior on OFAGE. Their mobility was also greatly changed after treatment with topoisomerase II, suggesting that the amplified DNA's were either supercoiled or concatenated circles.
D002460:Cell Line; D002875:Chromosomes; D004247:DNA / Q000235:genetics*; D004352:Drug Resistance, Microbial; D004586:Electrophoresis; D005111:Extrachromosomal Inheritance; D005784:Gene Amplification / Q000187:drug effects; D007895:Leishmania tropica / Q000187:drug effects / Q000235:genetics*; D008727:Methotrexate / Q000494:pharmacology; D009693:Nucleic Acid Hybridization
D004247:DNA; D008727:Methotrexate