title: Knockout of the abundant Trichomonas vaginalis hydrogenosomal membrane protein TvHMP23 increases hydrogenosome size but induces no compensatory up-regulation of paralogous copies.
abstract: The Trichomonas vaginalis genome encodes up to 60000 genes, many of which stem from genome duplication events. Paralogous copies thus accompany most T. vaginalis genes, a phenomenon that limits genetic manipulation. We characterized one of the parasite's most abundant hydrogenosomal membrane proteins, TvHMP23, which is phylogenetically distinct from canonical metabolite carriers, and which localizes to the inner hydrogenosomal membrane as shown through sub-organellar fractionation and protease protection assays. Knockout of Tvhmp23 through insertion of the selectable neomycin marker led to a size increase of hydrogenosomes, the first knockout-induced phenotypes reported for Trichomonas, but no growth impairment. The transcriptional response of its four paralogous copies then analyzed revealed that they are not up-regulated, and hence do not compensate for the Tvhmp23 knockout.
authors.affiliation: Institute of Molecular Evolution, Heinrich-Heine-Universität Düsseldorf, D-40225 Düsseldorf, Germany.
authors.forename: Xavier Pereira
authors.initials: XP
authors.lastname: Brás
authors.forename: Verena
authors.initials: V
authors.lastname: Zimorski
authors.forename: Kathrin
authors.initials: K
authors.lastname: Bolte
authors.forename: Uwe-G
authors.initials: UG
authors.lastname: Maier
authors.forename: William F
authors.initials: WF
authors.lastname: Martin
authors.forename: Sven B
authors.initials: SB
authors.lastname: Gould
mesh_terms: D055786:Gene Knockout Techniques; D059765:Homologous Recombination; D007425:Intracellular Membranes / Q000378:metabolism*; D008565:Membrane Proteins / Q000172:deficiency / Q000235:genetics* / Q000378:metabolism; D053141:Organelle Size; D010641:Phenotype; D021381:Protein Transport; D015800:Protozoan Proteins / Q000235:genetics* / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid; D014246:Trichomonas vaginalis / Q000166:cytology / Q000235:genetics*; D015854:Up-Regulation
chemical_list: D008565:Membrane Proteins; D015800:Protozoan Proteins