title: Evidence that transport of iron from the lysosome to the cytosol in African trypanosomes is mediated by a mucolipin orthologue.
abstract: Bloodstream-form Trypanosoma brucei acquire iron by receptor-mediated endocytosis of host transferrin. However, the mechanism(s) by which iron is then transferred from the lysosome to the cytosol are unresolved. Here, we provide evidence for the involvement of a protein (TbMLP) orthologous to the mammalian endolysosomal cation channel Mucolipin 1. In T. brucei, we show that this protein is localized to the single parasite lysosome. TbMLP null mutants could only be generated in the presence of an expressed ectopic copy, suggesting that the protein is essential. RNAi-mediated ablation resulted in a growth defect in vitro and led to a sevenfold increase in susceptibility to the iron-chelators deferoxamine and salicylhydroxamic acid. Conditional null mutants remained viable when the ectopic copy was repressed, but were hypersensitive to deferoxamine and displayed a growth defect similar to that observed following RNAi. The conditional nulls also retained virulence in vivo in the absence of the doxycycline inducer. These data provide strong evidence that TbMLP has a role in import of iron into the cytosol of African trypanosomes. They also indicate that even when expression is greatly reduced, there is sufficient protein, or an alternative mechanism, to provide the parasite with an adequate supply of cytosolic iron.
authors.affiliation: Department of Pathogen Molecular Biology, London School of Hygiene and Tropical Medicine, Keppel St., London, WC1E 7HT, UK. martin.taylor@lshtm.ac.uk
authors.forename: Martin C
authors.initials: MC
authors.lastname: Taylor
authors.forename: Alex P
authors.initials: AP
authors.lastname: McLatchie
authors.forename: John M
authors.initials: JM
authors.lastname: Kelly
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D003600:Cytosol / Q000737:chemistry*; D003676:Deferoxamine / Q000494:pharmacology; D005260:Female; D017353:Gene Deletion; D007501:Iron / Q000378:metabolism*; D008247:Lysosomes / Q000737:chemistry*; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism*; D034622:RNA Interference; D012457:Salicylamides / Q000494:pharmacology; D050051:Transient Receptor Potential Channels / Q000235:genetics / Q000378:metabolism*; D014346:Trypanosoma brucei brucei / Q000235:genetics / Q000378:metabolism*
chemical_list: D015800:Protozoan Proteins; D012457:Salicylamides; D050051:Transient Receptor Potential Channels; C005703:salicylhydroxamic acid; D007501:Iron; D003676:Deferoxamine