Molecular characterization of the MRPA transporter and antimony uptake in four New World Leishmania spp. susceptible and resistant to antimony

Moreira, Douglas S.; Monte-Neto, Rubens L.; Andrade, Juvana Moreira; Santi, Ana Maria Murta; Reis, Priscila G.; Frézard, Frédéric; Murta, Silvane Maria Fonseca

doi:10.1016/j.ijpddr.2013.08.001

Cited by 33 publications

(51 citation statements)

References 63 publications

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“…AQP1 has been shown to import Sb(III) and As(III) in Leishmania [4] and the decreased Sb(III) uptake due to reduced expression of AQP1 was associated with trivalent antimony resistance [4][5][6]. The copy number of the AQP1 gene derived from the polyploid chromosome 31 was decreased in Leishmania major antimony-resistant mutants [7] and the AQP1 gene was deleted in Sb(III) L. guyanensis resistant mutants [8].…”

Section: Leishmaniamentioning

confidence: 99%

Generation of an aquaglyceroporin AQP1 null mutant in Leishmania major

Plourde

Ubeda

Mandal

et al. 2015

Molecular and Biochemical Parasitology

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Section: Leishmaniamentioning

confidence: 99%

Generation of an aquaglyceroporin AQP1 null mutant in Leishmania major

Plourde

Ubeda

Mandal

et al. 2015

Molecular and Biochemical Parasitology

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“…One of the most characteristic mechanisms of antimony resistance in Leishmania is drug efflux mediated by ABC (ATP-binding cassette) transporters such as MRPA (formerly PGPA)/ABCC3 (3,4) or ABCI4 (5), which results in a reduced degree of antimony accumulation in parasites. ABC transporters are one of the largest protein families known; they are highly evolutionarily conserved from bacteria to humans and are involved in the transport of different compounds through biological membranes.…”

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confidence: 99%

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

Perea

Manzano

Castanys

et al. 2016

Antimicrob Agents Chemother

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Treatment for leishmaniasis, which is caused by Leishmania protozoan 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 in Leishmania is 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 in Leishmania have been associated with drug resistance. In this study, we have focused our interest to characterize LABCG2's involvement in drug resistance in Leishmania. Leishmania major parasites 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 that Leishmania parasites overexpressing LABCG2 were resistant to antimony, as they demonstrated a reduced accumulation of Sb III due to an increase in drug efflux. Additionally, LABCG2 was able to transport thiols in the presence of Sb III . 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, Leishmania LABCG2 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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“…AQP1 from Leishmania, the closest homologue of human AQP9, is permeated by water or small uncharged solutes, and nitrate ions probably do not pass through AQP1. In aqueous solution at physiological pH, trivalent antimoniate (Sb III ) exists mainly in the trihydroxylated uncharged form, Sb(OH) 3 , which structurally resembles glycerol (12 (30). Interestingly, both of these Sb III and nitrate combinations are very toxic for these resistant parasites.…”

Section: Discussionmentioning

confidence: 99%

“…The intracellular accumulation of antimony was measured as previously reported (30). Briefly, log-phase Leishmania promastigotes were washed twice with HEPES-glucose (HG) buffer (20 mM HEPES, 0.15 M NaCl, 10 mM glucose, pH 7.2) and resuspended in this buffer at a density of 1.0 ϫ 10 8 cells/ml.…”

Section: Methodsmentioning

confidence: 99%

Silver and Nitrate Oppositely Modulate Antimony Susceptibility through Aquaglyceroporin 1 in Leishmania (Viannia) Species

Andrade

Baba

Machado-de-Ávila

et al. 2016

Antimicrob Agents Chemother

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d Antimony (Sb) resistance in leishmaniasis chemotherapy has become one of the major challenges to the control of this spreading worldwide public health problem. Since the plasma membrane pore-forming protein aquaglyceroporin 1 (AQP1) is the major route of Sb uptake in Leishmania, functional studies are relevant to characterize drug transport pathways in the parasite. We generated AQP1-overexpressing Leishmania guyanensis and L. braziliensis mutants and investigated their susceptibility to the trivalent form of Sb (Sb III ) in the presence of silver and nitrate salts. Both AQP1-overexpressing lines presented 3-to 4-fold increased AQP1 expression levels compared with those of their untransfected counterparts, leading to an increased Sb III susceptibility of about 2-fold. Competition assays using silver nitrate, silver sulfadiazine, or silver acetate prior to Sb III exposure increased parasite growth, especially in AQP1-overexpressing mutants. Surprisingly, Sb III -sodium nitrate or Sb III -potassium nitrate combinations showed significantly enhanced antileishmanial activities compared to those of Sb III alone, especially against AQP1-overexpressing mutants, suggesting a putative nitrate-dependent modulation of AQP1 activity. The intracellular level of antimony quantified by graphite furnace atomic absorption spectrometry showed that the concomitant exposure to Sb III and nitrate favors antimony accumulation in the parasite, increasing the toxicity of the drug and culminating with parasite death. This is the first report showing evidence of AQP1-mediated Sb III susceptibility modulation by silver in Leishmania and suggests the potential antileishmanial activity of the combination of nitrate salts and Sb III .

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Molecular characterization of the MRPA transporter and antimony uptake in four New World Leishmania spp. susceptible and resistant to antimony

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Cited by 33 publications

References 63 publications

Generation of an aquaglyceroporin AQP1 null mutant in Leishmania major

Generation of an aquaglyceroporin AQP1 null mutant in Leishmania major

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

Silver and Nitrate Oppositely Modulate Antimony Susceptibility through Aquaglyceroporin 1 in Leishmania (Viannia) Species

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