Verner Zdeněk, Cermáková Petra, Skodová Ingrid, Kriegová Eva, Horváth Anton, Lukes Julius
Biology Centre, Institute of Parasitology, Czech Academy of Sciences, Ceské Budejovice Budweis, Czech Republic.
Mol Biochem Parasitol. 2011 Feb;175(2):196-200. doi: 10.1016/j.molbiopara.2010.11.003. Epub 2010 Nov 11.
The requirement of complex I (NADH:ubiquionone oxidoreductase) for respiration in Trypanosoma brucei is controversial. Recent identification of homologues of its subunits in mitochondrial proteome resolved a question of its presence or absence. However, with one exception, no data have been available concerning the function(s) of complex I or its subunits. Here we present a functional RNAi study of three (NUBM, NUKM, NUEM) putative subunits of this complex. Although no changes were detected in growth, mitochondrial membrane potential or reactive oxygen species production in cell lines depleted for target transcript, the NUBM and NUKM RNAi knock-downs showed decreased specific NADH:ubiquinone oxidoreductase activity. Moreover, glycerol gradients of all cell lines revealed the presence of two distinct peaks of NADH dehydrogenase activity, with shifted sensitivity to inhibitors of complex I upon RNAi induction. Thus complex I is not only present in the procyclic stage of T. brucei 29-13 strain, but it does participate in electron transport chain.
布氏锥虫呼吸作用对复合体I(NADH:泛醌氧化还原酶)的需求存在争议。近期在线粒体蛋白质组中鉴定出其亚基的同源物,解决了其是否存在的问题。然而,除了一个例外,关于复合体I及其亚基的功能尚无数据。在此,我们对该复合体的三个假定亚基(NUBM、NUKM、NUEM)进行了功能性RNA干扰研究。尽管在耗尽靶转录本的细胞系中,未检测到生长、线粒体膜电位或活性氧产生的变化,但NUBM和NUKM的RNA干扰敲低显示特定的NADH:泛醌氧化还原酶活性降低。此外,所有细胞系的甘油梯度显示存在两个不同的NADH脱氢酶活性峰,RNA干扰诱导后对复合体I抑制剂的敏感性发生了变化。因此,复合体I不仅存在于布氏锥虫29-13株的前循环期,而且确实参与电子传递链。