Modulation of gene expression in drug resistant Leishmania is associated with gene amplification, gene deletion and chromosome aneuploidy

Ubeda, Jean-Michel; Légaré, Danielle; Raymond, Frédéric; Ouameur, Amin Ahmed; Boisvert, Sébastien; Rigault, Philippe; Corbeil, Jacques; Tremblay, Michel J.; Olivier, Martin; Papadopoulou, Barbara; Ouellette, Marc

doi:10.1186/gb-2008-9-7-r115

Cited by 151 publications

(220 citation statements)

References 89 publications

(101 reference statements)

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“…A copy of the targeted gene was always present as a result of chromosome duplications (tri-or tetrasomy), wholegenome duplication, or translocation of the chromosome fragment carrying the target gene. Aneuploidy was also observed in laboratory strains of L. major and L. infantum after induction of drug resistance: transcriptional profiling using microarrays identified altered expression of genes on six different chromosomes in both species [10,11]. Quantitative Southern blot and comparative genomic hybridizations confirmed that the variation in expression corresponded to changes in the copy number of these chromosomes.…”

Section: The Flexible Genomes Of Leishmaniamentioning

confidence: 84%

“…By contrast, chromosome gain or loss (aneuploidy) in multicellular eukaryotes is either lethal or results in severe abnormalities, for example trisomy of chromosome 21 in humans with Down syndrome [6]. Despite the predominantly negative effects of aneuploidy, supernumerary chromosomes have been reported in tumor cells, yeast, and recently the protozoan Leishmania [7][8][9][10][11].…”

Section: The Flexible Genomes Of Leishmaniamentioning

confidence: 99%

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Adaptive mechanisms in pathogens: universal aneuploidy in Leishmania

Mannaert

Downing

Imamura

et al. 2012

Trends in Parasitology

129

102

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Section: The Flexible Genomes Of Leishmaniamentioning

confidence: 84%

Section: The Flexible Genomes Of Leishmaniamentioning

confidence: 99%

Adaptive mechanisms in pathogens: universal aneuploidy in Leishmania

Mannaert

Downing

Imamura

et al. 2012

Trends in Parasitology

129

102

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“…3A). Reduced RNA levels in Leishmania are usually due to a decrease in gene copy number (61,62), although other mechanisms may also occur (63). Interestingly, in the yeast S. cerevisiae that also transports AdoMet, resistance to SNF was shown to correlate with loss of function mutations in the AdoMet transporter Sam3 (20).…”

Section: Discussionmentioning

confidence: 99%

High Affinity S-Adenosylmethionine Plasma Membrane Transporter of Leishmania Is a Member of the Folate Biopterin Transporter (FBT) Family

Dridi¹,

Ouameur²,

Ouellette

2010

Journal of Biological Chemistry

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S-Adenosylmethionine (AdoMet) is an important methyl group donor that plays a central role in many essential biochemical processes. The parasite Leishmania can both synthesize and transport AdoMet. Leishmania cells resistant to the antifolate methotrexate due to a rearrangement in folate biopterin transporter (FBT) genes were cross-resistant to sinefungin, an AdoMet analogue. FBT gene rearrangements were also observed in Leishmania major cells selected for sinefungin resistance. One of the rearranged FBT genes corresponded to the main AdoMet transporter (AdoMetT1) of Leishmania as determined by gene transfection and gene inactivation experiments. AdoMetT1 was determined to be a high affinity plasma membrane transporter expressed constitutively throughout the growth phases of the parasite. Leishmania cells selected for resistance or naturally insensitive to sinefungin had lower expression of AdoMetT1. A new function in one carbon metabolism, also a pathway of interest for chemotherapeutic interventions, is described for a novel class of membrane proteins found in diverse organisms.The parasite Leishmania is distributed globally and causes a variety of clinical symptoms ranging from self-healing cutaneous lesions to visceral infections that are usually fatal if left untreated (1, 2). Current first-line chemotherapy against leishmaniasis relies on a rather limited arsenal of drugs, including pentavalent antimonials, liposomal amphotericin B, or miltefosine (2). These drugs are associated with side effects, high costs, and drug resistance, and therefore, the search for new drugs and targets to control this parasite is warranted (3, 4).S-Adenosylmethionine (AdoMet or SAM) 5 is an important biological sulfonium compound recognized as the universal methyl donor for methylation of lipids, proteins, nucleic acids, and xenobiotics in living cells (5). AdoMet is also a donor of propylamine groups for the synthesis of polyamines and participates in the reverse trans-sulfuration pathway where AdoMet is converted into cysteine and glutathione and in Leishmania in the spermidine-glutathione conjugate trypanothione (6, 7). AdoMet is also used as a source of methylene groups, amino groups, and ribosyl groups in the synthesis of fatty acids, biotin, and the modified nucleoside epoxyqueusine of tRNAs (reviewed in Ref. 8).Most cells are capable of synthesizing AdoMet from L-methionine and ATP, in a process requiring the enzyme methionine adenosyltransferase (MAT). The gene coding for this enzyme is present in most sequenced organisms (reviewed in Ref. 9). However, in some Rickettsia strains, the MAT gene is inactivated by a mutation (10), and in the fungus Pneumocystis carinii, no MAT activity has been detected (11). These organisms have the rare distinction of meeting their AdoMet needs by importing it (12). The Rickettsia transporter is part of the drug/ metabolite transporter superfamily (10), and the identity of the Pneumocystis transporter is not known. Transport of AdoMet across the plasma membrane does not occur to a signifi...

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“…Although methotrexate (MTX) is not used against Leishmania, it is nonetheless very active and has been extensively studied. Leishmania cells selected for MTX resistance undergo amplification of the dihydrofolate reductasethymidylate synthase (DHFR-TS) and pteridine reductase 1 (PTR1) genes, which encode primary (30,31) and secondary targets of MTX (32,33), respectively. Because of this unique combination of drug target relationship with MTX, we first validated Cos-Seq with parasites selected under MTX pressure.…”

Section: Significancementioning

confidence: 99%

“…A recent study has highlighted the role of protein phosphatase 2A in the mode of MTX action in mammalian cells (34). Thus, considering that MTX has similar mechanisms of action and entails comparable resistance mechanisms in Leishmania and mammalian cells (30,31,35), the two phosphatase-related genes emerged as reasonable candidates. Indeed, on the transfection of LinJ.34.2310 and LinJ.…”

Section: Significancementioning

confidence: 99%

Cos-Seq for high-throughput identification of drug target and resistance mechanisms in the protozoan parasite Leishmania

Gazanion

Fernández-Prada

Papadopoulou

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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106

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Innovative strategies are needed to accelerate the identification of antimicrobial drug targets and resistance mechanisms. Here we develop a sensitive method, which we term Cosmid Sequencing (or "Cos-Seq"), based on functional cloning coupled to next-generation sequencing. Cos-Seq identified >60 loci in the Leishmania genome that were enriched via drug selection with methotrexate and five major antileishmanials (antimony, miltefosine, paromomycin, amphotericin B, and pentamidine). Functional validation highlighted both known and previously unidentified drug targets and resistance genes, including novel roles for phosphatases in resistance to methotrexate and antimony, for ergosterol and phospholipid metabolism genes in resistance to miltefosine, and for hypothetical proteins in resistance to paromomycin, amphothericin B, and pentamidine. Several genes/loci were also found to confer resistance to two or more antileishmanials. This screening method will expedite the discovery of drug targets and resistance mechanisms and is easily adaptable to other microorganisms.

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Modulation of gene expression in drug resistant Leishmania is associated with gene amplification, gene deletion and chromosome aneuploidy

Cited by 151 publications

References 89 publications

Adaptive mechanisms in pathogens: universal aneuploidy in Leishmania

Adaptive mechanisms in pathogens: universal aneuploidy in Leishmania

High Affinity S-Adenosylmethionine Plasma Membrane Transporter of Leishmania Is a Member of the Folate Biopterin Transporter (FBT) Family

Cos-Seq for high-throughput identification of drug target and resistance mechanisms in the protozoan parasite Leishmania

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