scholarly journals Characterization of an ABCG-Like Transporter from the Protozoan Parasite Leishmania with a Role in Drug Resistance and Transbilayer Lipid Movement

2008 ◽  
Vol 52 (10) ◽  
pp. 3573-3579 ◽  
Author(s):  
Esther Castanys-Muñoz ◽  
José María Pérez-Victoria ◽  
Francisco Gamarro ◽  
Santiago Castanys
Keyword(s):  
Drug Resistance ◽  
Plasma Membrane ◽  
Treatment Failure ◽  
Tumor Cells ◽  
Abc Transporters ◽  
Protozoan Parasite ◽  
Protozoan Parasites ◽  
Resistance Phenotype ◽  
Parasite Leishmania

ABSTRACT Leishmaniasis treatment is hampered by the increased appearance of treatment failure. ATP-binding cassette (ABC) transporters are usually involved in drug resistance both in tumor cells and in microorganisms. Here we report the characterization of an ABCG-like transporter, LiABCG6, localized mainly at the plasma membrane in Leishmania protozoan parasites. When overexpressed, this half-transporter confers significant resistance to the leishmanicidal agents miltefosine and sitamaquine. This resistance phenotype is mediated by a reduction in intracellular drug accumulation. LiABCG6 also reduces the accumulation of short-chain fluorescent phospholipid analogues of phosphatidylcholine, phosphatidylethanolamine, and phosphatidylserine. As a whole, these results suggest that LiABCG6 could be implicated in phospholipid trafficking and drug resistance.

scholarly journals PDR Transporter ABC1 Is Involved in the Innate Azole Resistance of the Human Fungal Pathogen Fusarium keratoplasticum

2021 ◽  
Vol 12 ◽  
Author(s):  
Jasper Elvin James ◽  
Erwin Lamping ◽  
Jacinta Santhanam ◽  
Richard David Cannon
Keyword(s):  
Drug Resistance ◽  
Abc Transporters ◽  
Plant Pathogens ◽  
Efflux Pump ◽  
Logarithmic Phase ◽  
Azole Resistance ◽  
Mrna Levels ◽  
Multidrug Efflux Pump ◽  
Resistance Phenotype ◽  
Azole Antifungals

Fusarium keratoplasticum is arguably the most common Fusarium solani species complex (FSSC) species associated with human infections. Invasive fusariosis is a life-threatening fungal infection that is difficult to treat with conventional azole antifungals. Azole drug resistance is often caused by the increased expression of pleiotropic drug resistance (PDR) ATP-binding cassette (ABC) transporters of the ABCG sub-family. Most investigations of Fusarium ABC transporters associated with azole antifungal drug resistance are limited to plant pathogens. Through the manual curation of the entire ABCG protein family of four FSSC species including the fully annotated genome of the plant pathogen Nectria haematococca we identified PDR transporters ABC1 and ABC2 as the efflux pump candidates most likely to be associated with the innate azole resistance phenotype of Fusarium keratoplasticum. An initial investigation of the transcriptional response of logarithmic phase F. keratoplasticum cells to 16 mg/L voriconazole confirmed strong upregulation (372-fold) of ABC1 while ABC2 mRNA levels were unaffected by voriconazole exposure over a 4 h time-period. Overexpression of F. keratoplasticum ABC1 and ABC2 in the genetically modified Saccharomyces cerevisiae host ADΔΔ caused up to ∼1,024-fold increased resistance to a number of xenobiotics, including azole antifungals. Although ABC1 and ABC2 were only moderately (20% and 10%, respectively) expressed compared to the Candida albicans multidrug efflux pump CDR1, overexpression of F. keratoplasticum ABC1 caused even higher resistance levels to certain xenobiotics (e.g., rhodamine 6G and nigericin) than CDR1. Our investigations suggest an important role for ABC1 orthologues in the innate azole resistance phenotype of FSSC species.

scholarly journals ABC transporters in the protozoan parasite Leishmania

2001 ◽  
Vol 4 (3) ◽  
pp. 159-166 ◽  
Author(s):  
José M. Pérez-Victoria ◽  
Adriana Parodi-Talice ◽  
Cristina Torres ◽  
Francisco Gamarro ◽  
Santiago Castanys
Keyword(s):  
Abc Transporters ◽  
Protozoan Parasite ◽  
Parasite Leishmania

scholarly journals Characterization of Treatment Failure in Efficacy Trials of Drugs against Plasmodium vivax by Genotyping Neutral and Drug Resistance-Associated Markers

2011 ◽  
Vol 55 (9) ◽  
pp. 4479-4481 ◽  
Author(s):  
Celine Barnadas ◽  
Cristian Koepfli ◽  
Harin A. Karunajeewa ◽  
Peter M. Siba ◽  
Timothy M. E. Davis ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Treatment Failure ◽  
Plasmodium Vivax ◽  
Single Nucleotide ◽  
Content Type ◽  
Molecular Approaches ◽  
Efficacy Trials ◽  
Intervention Trials ◽  
Single Nucleotide Polymorphism Typing

ABSTRACTPlasmodium vivaxintervention trials customarily report uncorrected treatment failure rates. Application of recrudescence-reinfection genotyping and drug resistance single-nucleotide polymorphism typing to a 4-arm comparative efficacy trial illustrated that molecular approaches can assist in understanding the relative contributions of true drug resistance (recurrent with same genotype) and new infections to treatment failure. The PCR-corrected adequate clinical and parasitologic response may constitute an informative secondary endpoint in futureP. vivaxdrug trials.

Purification and biochemical characterization of NpABCG5/NpPDR5, a plant pleiotropic drug resistance transporter expressed in Nicotiana tabacum BY-2 suspension cells

2017 ◽  
Vol 474 (10) ◽  
pp. 1689-1703 ◽  
Author(s):  
Frédéric Toussaint ◽  
Baptiste Pierman ◽  
Aurélie Bertin ◽  
Daniel Lévy ◽  
Marc Boutry
Keyword(s):  
Electron Microscopy ◽  
Drug Resistance ◽  
Nicotiana Tabacum ◽  
Abc Transporters ◽  
Plasma Membranes ◽  
Biochemical Characterization ◽  
Specific Activity ◽  
Pleiotropic Drug Resistance ◽  
Suspension Cells

Pleiotropic drug resistance (PDR) transporters belong to the ABCG subfamily of ATP-binding cassette (ABC) transporters and are involved in the transport of various molecules across plasma membranes. During evolution, PDR genes appeared independently in fungi and in plants from a duplication of a half-size ABC gene. The enzymatic properties of purified PDR transporters from yeast have been characterized. This is not the case for any plant PDR transporter, or, incidentally, for any purified plant ABC transporter. Yet, plant PDR transporters play important roles in plant physiology such as hormone signaling or resistance to pathogens or herbivores. Here, we describe the expression, purification, enzymatic characterization and 2D analysis by electron microscopy of NpABCG5/NpPDR5 from Nicotiana plumbaginifolia, which has been shown to be involved in the plant defense against herbivores. We constitutively expressed NpABCG5/NpPDR5, provided with a His-tag in a homologous system: suspension cells from Nicotiana tabacum (Bright Yellow 2 line). NpABCG5/NpPDR5 was targeted to the plasma membrane and was solubilized by dodecyl maltoside and purified by Ni-affinity chromatography. The ATP-hydrolyzing specific activity (27 nmol min−1 mg−1) was stimulated seven-fold in the presence of 0.1% asolectin. Electron microscopy analysis indicated that NpABCG5/NpPDR5 is monomeric and with dimensions shorter than those of known ABC transporters. Enzymatic data (optimal pH and sensitivity to inhibitors) confirmed that plant and fungal PDR transporters have different properties. These data also show that N. tabacum suspension cells are a convenient host for the purification and biochemical characterization of ABC transporters.

scholarly journals The LABCG2 Transporter from the Protozoan Parasite Leishmania Is Involved in Antimony Resistance

2016 ◽  
Vol 60 (6) ◽  
pp. 3489-3496 ◽  
Author(s):  
Ana Perea ◽  
José Ignacio Manzano ◽  
Santiago Castanys ◽  
Francisco Gamarro
Keyword(s):  
Drug Resistance ◽  
Plasma Membrane ◽  
Drug Targets ◽  
Molecular Mechanisms ◽  
Leishmania Major ◽  
Fluorescent Protein ◽  
Protozoan Parasite ◽  
Content Type ◽  
Northern India ◽  
Antimony Resistance

Treatment for leishmaniasis, which is caused byLeishmaniaprotozoan parasites, currently relies on a reduced arsenal of drugs. However, the significant increase in the incidence of drug therapeutic failure and the growing resistance to first-line drugs like antimonials in some areas of Northern India and Nepal limit the control of this parasitic disease. Understanding the molecular mechanisms of resistance inLeishmaniais now a matter of urgency to optimize drugs used and to identify novel drug targets to block or reverse resistant mechanisms. Some members of the family of ATP-binding cassette (ABC) transporters inLeishmaniahave been associated with drug resistance. In this study, we have focused our interest to characterize LABCG2's involvement in drug resistance inLeishmania. Leishmania majorparasites overexpressing the ABC protein transporter LABCG2 were generated in order to assess how LABCG2 is involved in drug resistance. Assays of susceptibility to different leishmanicidal agents were carried out. Analysis of the drug resistance profile revealed thatLeishmaniaparasites overexpressing LABCG2 were resistant to antimony, as they demonstrated a reduced accumulation of SbIIIdue to an increase in drug efflux. Additionally, LABCG2 was able to transport thiols in the presence of SbIII. Biotinylation assays using parasites expressing LABCG2 fused with an N-terminal green fluorescent protein tag revealed that LABCG2 is partially localized in the plasma membrane; this supports data from previous studies which suggested that LABCG2 is localized in intracellular vesicles that fuse with the plasma membrane during exocytosis. In conclusion,LeishmaniaLABCG2 probably confers antimony resistance by sequestering metal-thiol conjugates within vesicles and through further exocytosis by means of the parasite's flagellar pocket.

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