Generation of an aquaglyceroporin AQP1 null mutant in Leishmania major

作者：Plourde Marie; Ubeda Jean Michel; Mandal Goutam; do Monte Neto Rubens Lima; Mukhopadhyay Rita; Ouellette Marc^*

来源：Molecular and Biochemical Parasitology, 2015, 201(2): 108-111.

DOI：10.1016/j.molbiopara.2015.07.003

• Leishmania
• Aquaglyceroporin AQPI
• Antimony resistance
• Transport assays
• Osmoregulation

The Leishmania aquaglyceroporin AQP1 plays an important physiological role in water and uncharged polar solutes transport, volume regulation, osmotaxis, and is a key determinant of antimony resistance. By targeted gene disruption, we generated a Leishmania major promastigote AQP1 null mutant. This required several attempts but a chromosomal null AQP1 mutant was obtained by loss of heterozygosity in the presence of a rescue plasmid encoding AQP1. Growth in the absence of selection led to the loss of the rescuing plasmid, indicating that AQP1 is not essential for Leishmania viability. The AQP1-null mutant was resistant to antimonyl tartrate (SbIII) and arsenite (AsIII) due to a decrease import of these metalloids. It also exhibited alterations in its osmoregulation abilities compared with wild-type cells. This is the first report of the generation of a genetic AQP1 null mutant in Leishmania parasite, confirming its physiological function and role in resistance to antimonials, the therapeutic mainstay against Leishmania.

• 出版日期2015-6