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Nucleic Acids Research, 1993, Vol. 21, No. 8 1895-1901
© 1993

MOLECULAR BIOLOGY

Homologous recombination between direct repeat sequences yields P-glycoprotein containing amplicons in arsenite resistant Leishmania

Katherine Grondin, Barbara Papadopoulon and Marc Ouellette*

Service d'Infectiologie du Centre de Recherche du CHUL et Département de Microbiologie, Faculté de Médecine Université Laval, Québec, Canada

* To whom correspondence should be addressed at: Service d'lnfectiologie du Centre de Recherche du CHUL, 2705 boul. Laurier, Ste-Foy, QuéTxc G1V 4G2, Canada

Received December 21, 1992. Revised March 17, 1993. Accepted March 17, 1993.

The protozoan parasite Leishmania often responds to drug pressure by amplifying part of its genome. At least two loci derived from the same 800 kb chromosome were amplified either as extrachromosomal circles or linear fragments after sodium arsenlte selection. A 50 kb linear amplicon was detected in six Independent arsenlte mutants and revertants grown in absence of arsenlte rapidly lost the amplicon and part of their resistance. The circular extrachromosomal ampllcons, all derived from the H locus of Leishmania, were characterized more extensively. In all cases, direct repeated sequences appeared to be Involved in the formation of circular ampllcons. Most amplicons were generated after homologous recombination between two linked P-glycoproteln genes. This recombination event was, in two cases, associated with the loss of one allele of the chromosomal copy. A novel rearrangement point was found in a mutant where the amplicon was created by recombination between two 541 bp direct repeats surrounding the P-glycoproteln gene present at the H locus. It Is also at one of these repeats that an H circle with large inverted duplications was formed. We propose that the presence of repeated sequences in the H locus facilitates the amplification of the drug resistance genes concentrated In this locus.


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