title: Drug uptake and modulation of drug resistance in Leishmania by an aquaglyceroporin.
abstract: Leishmaniasis is a protozoan parasitic disease that affects 12 million people worldwide. The first line choice for the treatment of this disease is antimonial drugs. In the endemic regions, resistance to this class of drugs is a major impediment to treatment. Microbes often become resistant to drugs by mutation or down-regulation of uptake systems, but the uptake system for the antimonial drugs in Leishmania is unknown. In other organisms, aquaglyceroporins have been shown to facilitate uptake of trivalent metalloids. In this study, we report the identification and characterization of aquaglyceroporins from Leishmania major (LmAQP1) and Leishmania tarentolae (LtAQP1), respectively. These Leishmania proteins have the conserved signature motifs of aquaglyceroporins. Transfection of LmAQP1 into three species of Leishmania, L. tarentolae, Leishmania infantum, and L. major, produced hypersensitivity to both As(III) and Sb(III) in all three strains. Increased production of LmAQP1 was detected by immunoblotting. Drug-resistant parasites with various mutations leading to resistance mechanisms became hypersensitive to both metalloids after expression of LmAQP1. Increased rates of uptake of As(III) or Sb(III) correlated with metalloid sensitivity of the wild type and drug-resistant transfectants. Transfection of LmAQP1 in a Pentostam-resistant field isolate also sensitized the parasite in the macrophage-associated amastigote form. One allele of LmAQP1 was disrupted in L. major, and the resulting cells became 10-fold more resistant to Sb(III). This is the first report of the uptake of a metalloid drug by an aquaglyceroporin in Leishmania, suggesting a strategy to reverse resistance in the field.
authors.affiliation: Department of Biochemistry and Molecular Biology, School of Medicine, Wayne State University, Detroit, Michigan 48201, USA.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Gourbal
authors.forename: Niluefer
authors.initials: N
authors.lastname: Sonuc
authors.forename: Hiranmoy
authors.initials: H
authors.lastname: Bhattacharjee
authors.forename: Danielle
authors.initials: D
authors.lastname: Legare
authors.forename: Shyam
authors.initials: S
authors.lastname: Sundar
authors.forename: Marc
authors.initials: M
authors.lastname: Ouellette
authors.forename: Barry P
authors.initials: BP
authors.lastname: Rosen
authors.forename: Rita
authors.initials: R
authors.lastname: Mukhopadhyay
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000981:Antiprotozoal Agents / Q000494:pharmacology; D020346:Aquaporins / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D001693:Biological Transport, Active; D003001:Cloning, Molecular; D016054:DNA, Protozoan / Q000235:genetics; D004351:Drug Resistance / Q000235:genetics; D006801:Humans; D007700:Kinetics; D007891:Leishmania / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism*; D007896:Leishmaniasis / Q000188:drug therapy / Q000469:parasitology; D008969:Molecular Sequence Data; D009154:Mutation; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid
chemical_list: D000981:Antiprotozoal Agents; D020346:Aquaporins; D016054:DNA, Protozoan; D015800:Protozoan Proteins