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Biochemistry |
t
pánka Va
á
ová1
Department of Parasitology, Faculty of Science, Charles University, Vini
ná 7, Prague, 128 44, Czech Republic1
Author for correspondence: Jan Tachezy. Tel: +420 2 21953207. Fax: +420 2 24919704. e-mail: tachezy{at}natur.cuni.cz
The main function of the hydrogenosome, a typical organelle of trichomonads, is to convert malate or pyruvate to H2, CO2 and acetate by a pathway associated with ATP synthesis. This pathway relies on activity of ironsulfur proteins such as pyruvate:ferredoxin oxidoreductase (PFOR), hydrogenase and ferredoxin. To examine the effect of iron availability on proper hydrogenosomal function, the metabolic activity of the hydrogenosome and expression of hydrogenosomal enzymes were compared in Tritrichomonas foetus maintained under iron-rich (150 µM iron nitrilotriacetate) or iron-restricted (180 µM 2,2-dipyridyl) conditions in vitro. The activities of PFOR and hydrogenase, and also production of acetate and H2, were markedly decreased or absent in iron-restricted trichomonads. Moreover, a decrease in activity of the hydrogenosomal malic enzyme, which is a non-FeS protein, was also observed. Impaired function of hydrogenosomes under iron-restricted conditions was compensated for by activation of the cytosolic pathway, mediating conversion of pyruvate to ethanol via acetaldehyde. This metabolic switch was fully reversible. Production of hydrogen by iron-restricted trichomonads was restored to the level of organisms grown under iron-rich conditions within 3 h after addition of 150 µM iron nitrilotriacetate. Protein analysis of purified hydrogenosomes from iron-restricted cells showed decreased levels of proteins corresponding to PFOR, malic enzyme and ferredoxin. Accordingly, these cells displayed decreased steady-state level and synthesis of mRNAs encoding PFOR and hydrogenosomal malic enzyme. These data demonstrate that iron is essential for function of the hydrogenosome, show its involvement in the expression of hydrogenosomal proteins and indicate the presence of iron-dependent control of gene transcription in Tt. foetus.
Keywords: Tritrichomonas foetus, iron, carbohydrate metabolism, hydrogenosome, transcription
Abbreviations: Fe-NTA, iron nitrilotriacetate; PFOR, pyruvate:ferredoxin oxidoreductase
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