Journal
MOLECULAR AND BIOCHEMICAL PARASITOLOGY
Volume 201, Issue 2, Pages 108-111Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.molbiopara.2015.07.003
Keywords
Leishmania; Aquaglyceroporin AQPI; Antimony resistance; Transport assays; Osmoregulation
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Funding
- CIHR [15501]
- Herbert Weirtheim College of Medicine start up funds
- Government of Canada DFAIT post-doctoral research fellowship
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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. (C) 2015 Elsevier B.V. All rights reserved.
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