title: Depletion of GIM5 causes cellular fragility, a decreased glycosome number, and reduced levels of ether-linked phospholipids in trypanosomes.
abstract: Microbody division in mammalian cells, trypanosomes, and yeast depends on the PEX11 microbody membrane proteins. The function of PEX11 is not understood, and the suggestion that it affects microbody (peroxisome) numbers in mammals and yeast, because it plays a role in beta-oxidation of fatty acids, is controversial. PEX11 and two PEX11-related proteins, GIM5A and GIM5B, are the predominant membrane proteins of the microbodies (glycosomes) of Trypanosoma brucei. The compartmentation of glycosomal enzymes is essential in trypanosomes. Deletion of the GIM5A gene from the form of the parasite that lives in the mammalian blood has no effect on trypanosome growth, but depletion of GIM5B on a gim5a null background causes death. We show here that procyclic trypanosomes, adapted for life in the Tsetse fly vector, survive without GIM5A and with very low levels of GIM5B. The depleted cells have fewer glycosomes than usual and are osmotically fragile, which is a novel observation for a microbody defect. Thus trypanosomes require both GIM5B and PEX11 for the maintenance of normal glycosome numbers. Procyclic cells lacking GIM5A, like mouse cells partially defective in PEX11, have fewer ether-linked phospholipids, even when GIM5B levels are not reduced. Metabolite measurements on GIM5A/B-depleted bloodstream form trypanosomes suggested a change in the flux through the glycolytic pathway. We conclude that PEX11 family proteins play important roles in determining microbody membrane structure, with secondary effects on a subset of microbody metabolic pathways.
authors.affiliation: Zentrum für Molekulare Biologie Heidelberg, Im Neuenheimer Feld 282, D-69120 Heidelberg, Germany. voncken2001@yahoo.de
authors.forename: Frank
authors.initials: F
authors.lastname: Voncken
authors.forename: Jaap J
authors.initials: JJ
authors.lastname: van Hellemond
authors.forename: Iris
authors.initials: I
authors.lastname: Pfisterer
authors.forename: Alexander
authors.initials: A
authors.lastname: Maier
authors.forename: Stephan
authors.initials: S
authors.lastname: Hillmer
authors.forename: Christine
authors.initials: C
authors.lastname: Clayton
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002455:Cell Division / Q000502:physiology*; D017931:DNA Primers; D004986:Ether / Q000378:metabolism*; D017353:Gene Deletion; D018503:Genome, Protozoan; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D015388:Organelles / Q000378:metabolism*; D010743:Phospholipids / Q000378:metabolism*; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism*; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D014346:Trypanosoma brucei brucei / Q000166:cytology / Q000235:genetics / Q000502:physiology*
chemical_list: D017931:DNA Primers; C478482:GIM5A protein, Trypanosoma brucei; C478483:GIM5B protein, Trypanosoma brucei; D008565:Membrane Proteins; D010743:Phospholipids; D015800:Protozoan Proteins; D004986:Ether