Intracellular location of the ABC transporter PRP1 related to pentamidine resistance in Leishmania major

Coelho, Adriano C.; Yamashiro-Kanashiro, Edite H.; Bastos, Sueli F.; Mortara, Renato Arruda; Cotrim, Paulo C.

doi:10.1016/j.molbiopara.2006.08.013

Cited by 20 publications

(10 citation statements)

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“…The involvement of the intracellular ABCC proteins MRPA/ ABCC3 and PRP1/ABCC7 in antimony resistance has already been reported (2,3,15). However, only MRPA/ABCC3 has been shown to confer resistance both to Sb(V) and to Sb(III) (5) and to be amplified in field isolates derived from patients unresponsive to antimonials (18).…”

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“…Cells transfected with the ABCC8-GFP fusion showed a punctuated fluorescence signal located near the posterior end of the parasite. We did not carry subcellular localization experiments for the MRPA/ABCC3 and the PRP1/ ABCC7 proteins, as they are already known to be intracellular proteins (3,15), nor for the ABCC9 protein, given the lack of antimony resistance following the overexpression of its gene (see below). The N-terminal and C-terminal GFP-tagged versions of ABCC5 localized to the same tubular compartment (results not shown), suggesting that the C-terminal GFP tagging strategy is probably not interfering with the localization of the fusion proteins.…”

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Intracellular Localization of the ABCC Proteins of Leishmania and Their Role in Resistance to Antimonials

Leprohon

Légaré

Ouellette

2009

Antimicrob Agents Chemother

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The ABCC subfamily of proteins is composed of nine members in Leishmania. We report that all of these proteins have an intracellular localization and that the overexpression of at least four members, ABCC3, ABCC4, ABCC5, and ABCC7, can confer resistance to antimonials, the first-line drug against Leishmania.The protozoan parasite Leishmania is responsible for a variety of clinical manifestations, ranging from mild cutaneous infections to life-threatening visceral diseases (12). Pentavalent antimony [Sb(V)] containing compounds such as sodium stibogluconate (Pentostam) and N-methylglucamine (Glucantime) remain the first-line drugs against all forms of Leishmania infections in developing countries (19), but their efficacies are threatened by resistant parasites in several regions where the disease is endemic (9,17,21). Our previous in vitro work on metal resistance in Leishmania led to the definition of a model of resistance involving proteins of the ATP-binding cassette (ABC) superfamily (20). ABC proteins form one of the largest families of transmembrane proteins and are characterized by the presence of the strongly conserved nucleotide-binding domain (NBD), which is composed of three major motifs. Along with the Walker A and B motifs found in many nucleotide-binding proteins (23), the NBD is composed of a characteristic ABC signature "C" motif located just upstream of the Walker B site (13). Eukaryotic ABC proteins can be divided into eight different subfamilies (ABCA to ABCH) on the basis of gene structure and NBD sequence homologies. A previous survey indicated the presence of 42 ABC protein-coding genes in the genomes of Leishmania major and Leishmania infantum (16), but the latest version of the L. infantum genome (GeneDB V3.0) revealed the presence of a new member of the ABC superfamily (LinJ24_V3.1510). This gene seems to be specific to L. infantum since it is absent from the genome of L. major (GeneDB V5.1) and is found as a pseudogene of low homology in the genome of Leishmania braziliensis (GeneDB V2.0). A phylogenetic analysis of the L. infantum ABC proteins revealed that LinJ24_V3.1510 is the most divergent member of the ABCC subfamily in Leishmania (results not shown) and has been named ABCC9. While the Walker A and B motifs are well conserved in the ABCC proteins of Leishmania, some conserved residues essential to the function of ABC proteins (7) are absent from the C motif of ABCC9 (Fig. 1), and the functionality of this protein in L. infantum still needs to be established.Resistance to metalloids in Leishmania requires multiple steps, where Sb(V) is reduced to the trivalent form Sb(III), and the latter is conjugated to trypanothione (TSH), a bisglutathione-spermidine conjugate (6), before being transported inside an intracellular detoxification organelle by the ABC transporter MRPA/ABCC3 (15) or being extruded outside the cell by an ATP-dependent efflux pump of unknown identity (4). As the active extrusion of metalloids conjugated to thiols is a feature of the ABCC subfamily (14, 25), we hypothe...

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Intracellular Localization of the ABCC Proteins of Leishmania and Their Role in Resistance to Antimonials

Leprohon

Légaré

Ouellette

2009

Antimicrob Agents Chemother

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“…PRP1 is part of the Leishmania ABCC subfamily, which also includes MRPA (ABCC3), a protein involved in antimony resistance in Leishmania (8,14). PRP1 and MRPA are intracellular proteins and are possibly associated with the tubulovesicular element that is linked to exo-and endocytosis pathways (4,11,18). We hypothesized that PRP1 would transport pentamidine into an intracellular organelle that would be exocytosed by the cell throughout the flagellar pocket (4).…”

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confidence: 99%

“…PRP1 and MRPA are intracellular proteins and are possibly associated with the tubulovesicular element that is linked to exo-and endocytosis pathways (4,11,18). We hypothesized that PRP1 would transport pentamidine into an intracellular organelle that would be exocytosed by the cell throughout the flagellar pocket (4). These data are derived from work with promastigotes, and in this study, we have tested whether PRP1 can confer resistance to pentamidine in the more relevant amastigote stage of the parasite.…”

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confidence: 99%