title: A functional BH3 domain in an aquaporin from Leishmania infantum.
abstract: Despite the absence of sequences showing significant similarity to any of the members of the Bcl-2 family of proteins in protozoa, experiments carried out in yeast or trypanosomatids have demonstrated that ectopic expression of some of these members alters their response to different death stimuli. Because the BH3 domain is the smallest common signature in all the proteins of this family of apoptosis regulators and also because they are essential for molecular interactions between antagonistic members, we looked for sequences with significant similarity to the BH3 motif in the Leishmania infantum genome. Among the top scoring ones, we found the MYLALQNLGDEV amino-acid stretch at the C terminus of a previously described aquaporin, now renamed as Li-BH3AQP. This motif is highly conserved in homologous proteins from other species of the Leishmania genus. The association of Li-BH3AQP with human Bcl-XL was demonstrated by both co-immunoprecipitation and yeast two-hybrid experiments. Ectopic expression of Li-BH3AQP reduced viability of HeLa cells and this deleterious effect was abrogated by the simultaneous overexpression of Bcl-XL. Although we were not able to demonstrate a reduction in parasite viability when the protein was overexpressed in Leishmania promastigotes, a prodeath effect could be observed when the parasites overexpressing Li-BH3AQP were treated with staurosporine or antimycin A. Surprisingly, these parasites were more resistant, compared with wild-type parasites, to hypotonic stress or nutrient deprivation. The prodeath activity was abolished upon replacement of two highly conserved amino acids in this BH3 domain. Taken together, these results point to Li-BH3AQP as the first non-enzymatic protein ever described in trypanosomatids that is involved in cell death.
authors.affiliation: Departamento de Biología de Sistemas, Universidad de Alcalá , Alcalá de Henares 28805, Spain.
authors.forename: C M
authors.initials: CM
authors.lastname: Genes
authors.affiliation: Departamento de Biología de Sistemas, Universidad de Alcalá , Alcalá de Henares 28805, Spain.
authors.forename: H
authors.initials: H
authors.lastname: de Lucio
authors.affiliation: Laboratory of aptamers, Departamento de Bioquímica-Investigación, IRYCIS-Hospital Ramón y Cajal , Madrid, Spain.
authors.forename: V M
authors.initials: VM
authors.lastname: González
authors.affiliation: Departamento de Ciencias Biomédicas, Universidad de Alcalá, Facultad de Medicina , Alcalá de Henares 28805, Spain.
authors.forename: P A
authors.initials: PA
authors.lastname: Sánchez-Murcia
authors.affiliation: Departamento de Biología de Sistemas, Universidad de Alcalá , Alcalá de Henares 28805, Spain.
authors.forename: E
authors.initials: E
authors.lastname: Rico
authors.affiliation: Departamento de Ciencias Biomédicas, Universidad de Alcalá, Facultad de Medicina , Alcalá de Henares 28805, Spain.
authors.forename: F
authors.initials: F
authors.lastname: Gago
authors.affiliation: Department of Biochemistry, University of Lausanne , 155 Chemin des Boveresses, Epalinges 1066, Switzerland.
authors.forename: N
authors.initials: N
authors.lastname: Fasel
authors.affiliation: Departamento de Biología de Sistemas, Universidad de Alcalá , Alcalá de Henares 28805, Spain.
authors.forename: A
authors.initials: A
authors.lastname: Jiménez-Ruiz