title: Generation of an aquaglyceroporin AQP1 null mutant in Leishmania major.
abstract: 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.
authors.affiliation: Centre de Recherche en Infectiologie du Centre de Recherche du CHU and Département de Microbiologie, Immunologie et Infectiologie, Faculté de Médecine, Université Laval, Québec, Québec, Canada.
authors.forename: Marie
authors.initials: M
authors.lastname: Plourde
authors.affiliation: Centre de Recherche en Infectiologie du Centre de Recherche du CHU and Département de Microbiologie, Immunologie et Infectiologie, Faculté de Médecine, Université Laval, Québec, Québec, Canada.
authors.forename: Jean-Michel
authors.initials: JM
authors.lastname: Ubeda
authors.affiliation: Molecular Microbiology and Infectious Diseases, Herbert Wertheim College of Medicine, Miami, USA.
authors.forename: Goutam
authors.initials: G
authors.lastname: Mandal
authors.affiliation: Centro de Pesquisas René Rachou, Fundação Oswaldo Cruz, Belo Horizonte, Minas Gerais, Brazil.
authors.forename: Rubens Lima do
authors.initials: RL
authors.lastname: Monte-Neto
authors.affiliation: Molecular Microbiology and Infectious Diseases, Herbert Wertheim College of Medicine, Miami, USA.
authors.forename: Rita
authors.initials: R
authors.lastname: Mukhopadhyay
authors.affiliation: Centre de Recherche en Infectiologie du Centre de Recherche du CHU and Département de Microbiologie, Immunologie et Infectiologie, Faculté de Médecine, Université Laval, Québec, Québec, Canada. Electronic address: marc.ouellette@crchul.ulaval.ca.
authors.forename: Marc
authors.initials: M
authors.lastname: Ouellette
mesh_terms: D000966:Antimony Potassium Tartrate / Q000633:toxicity; D051397:Aquaglyceroporins / Q000172:deficiency*; D018053:Arsenites / Q000633:toxicity; D002470:Cell Survival / Q000187:drug effects; D004351:Drug Resistance; D055786:Gene Knockout Techniques; D018320:Leishmania major / Q000187:drug effects / Q000235:genetics* / Q000502:physiology; D064587:Osmoregulation
chemical_list: D051397:Aquaglyceroporins; D018053:Arsenites; D000966:Antimony Potassium Tartrate
keywords: Antimony resistance; Aquaglyceroporin AQP1; Leishmania; Osmoregulation; Transport assays