title: Enigmatic presence of mitochondrial complex I in Trypanosoma brucei bloodstream forms.
abstract: The presence of mitochondrial respiratory complex I in the pathogenic bloodstream stages of Trypanosoma brucei has been vigorously debated: increased expression of mitochondrially encoded functional complex I mRNAs is countered by low levels of enzymatic activity that show marginal inhibition by the specific inhibitor rotenone. We now show that epitope-tagged versions of multiple complex I subunits assemble into α and β subcomplexes in the bloodstream stage and that these subcomplexes require the mitochondrial genome for their assembly. Despite the presence of these large (740- and 855-kDa) multisubunit complexes, the electron transport activity of complex I is not essential under experimental conditions since null mutants of two core genes (NUBM and NUKM) showed no growth defect in vitro or in mouse infection. Furthermore, the null mutants showed no decrease in NADH:ubiquinone oxidoreductase activity, suggesting that the observed activity is not contributed by complex I. This work conclusively shows that despite the synthesis and assembly of subunit proteins, the enzymatic function of the largest respiratory complex is neither significant nor important in the bloodstream stage. This situation appears to be in striking contrast to that for the other respiratory complexes in this parasite, where physical presence in a life-cycle stage always indicates functional significance.
authors.affiliation: Seattle Biomedical Research Institute, Seattle, Washington, USA.
authors.forename: Sachin
authors.initials: S
authors.lastname: Surve
authors.forename: Meredith
authors.initials: M
authors.lastname: Heestand
authors.forename: Brian
authors.initials: B
authors.lastname: Panicucci
authors.forename: Achim
authors.initials: A
authors.lastname: Schnaufer
authors.forename: Marilyn
authors.initials: M
authors.lastname: Parsons
mesh_terms: D000818:Animals; D004579:Electron Transport; D042967:Electron Transport Complex I / Q000378:metabolism*; D015200:Fluoroimmunoassay; D051379:Mice; D008807:Mice, Inbred BALB C; D008928:Mitochondria / Q000201:enzymology* / Q000378:metabolism; D010084:Oxidation-Reduction; D051381:Rats; D014162:Transfection; D014346:Trypanosoma brucei brucei / Q000201:enzymology* / Q000378:metabolism
chemical_list: D042967:Electron Transport Complex I