Knock-downs of iron-sulfur cluster assembly proteins IscS and IscU down-regulate the active mitochondrion of procyclic Trypanosoma brucei

被引:60
作者
Smid, Ondrej
Horakova, Eva
Vilimova, Vanda
Hrdy, Ivan
Cammack, Richard
Horvath, Anton
Lukes, Julius
Tachezy, Jan
机构
[1] Charles Univ Prague, Dept Parasitol, Fac Sci, CR-12844 Prague 2, Czech Republic
[2] Univ S Bohemia, Fac Biol, Ceske Budejovice 35007, Czech Republic
[3] Acad Sci Czech Republ, Inst Parasitol, Ceske Budejovice 35007, Czech Republic
[4] Comenius Univ, Fac Nat Sci, Bratislava 84215, Slovakia
[5] Kings Coll London, Dept Life Sci, London WC2R 2LS, England
关键词
D O I
10.1074/jbc.M513781200
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Transformation of the metabolically down-regulated mitochondrion of the mammalian bloodstream stage of Trypanosoma brucei to the ATP-producing mitochondrion of the insect procyclic stage is accompanied by the de novo synthesis of citric acid cycle enzymes and components of the respiratory chain. Because these metabolic pathways contain multiple iron-sulfur (FeS) proteins, their synthesis, including the formation of FeS clusters, is required. However, nothing is known about FeS cluster biogenesis in trypanosomes, organisms that are evolutionarily distant from yeast and humans. Here we demonstrate that two mitochondrial proteins, the cysteine desulfurase TbiscS and the metallochaperone TbiscU, are functionally conserved in trypanosomes and essential for this parasite. Knock-downs of TbiscS and TbiscU in the procyclic stage by means of RNA interference resulted in reduced activity of the marker FeS enzyme aconitase in both the mitochondrion and cytosol because of the lack of FeS clusters. Moreover, down-regulation of TbiscS and TbiscU affected the metabolism of procyclic T. brucei so that their mitochondria resembled the organelle of the bloodstream stage; mitochondrial ATP production was impaired, the activity of the respiratory chain protein complex ubiquinol-cytochrome-c reductase was reduced, and the production of pyruvate as an end product of glucose metabolism was enhanced. These results indicate that mitochondrial FeS cluster assembly is indispensable for completion of the T. brucei life cycle.
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页码:28679 / 28686
页数:8
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