Brown B S V, Chi T B, Williams N
253 Biomedical Research Building, Department of Microbiology, State University of New York at Buffalo, 14214, Buffalo, NY, USA.
Mol Biochem Parasitol. 2001 Jul;115(2):177-87. doi: 10.1016/s0166-6851(01)00282-1.
The mitochondrial ATP synthase is developmentally regulated throughout the life cycle of the Trypanosoma brucei. The alpha and beta subunits of the F(1) moiety, and subunit 9 of the F(0) moiety of the T. brucei ATP synthase have been previously cloned and characterized. Here we have determined the chromosomal localization and developmental regulation of these three key subunits of the complex. Southern blot analysis indicates that all three of these genes are present as single copies in the T. brucei genome. Pulsed field gel analysis demonstrates that these genes are encoded in different chromosomes, and are thus not part of the same gene cluster. A comparison between the protein and steady state transcript levels for these subunits suggests that regulation of expression occurs predominantly posttranscriptionally. Comparison of mRNA stability for procyclic and bloodstream forms shows that the half life of the three transcripts is much shorter in bloodstream forms. The differences in transcript stability in the procyclic form for subunit 9 is greater than that for alpha and beta subunits, while the differences at the protein levels are comparable. These results suggest that there may be further posttranscriptional regulation of subunit 9.
线粒体ATP合酶在布氏锥虫的整个生命周期中受到发育调控。布氏锥虫ATP合酶F1部分的α和β亚基以及F0部分的9号亚基先前已被克隆和鉴定。在此,我们确定了该复合体这三个关键亚基的染色体定位和发育调控。Southern印迹分析表明,这三个基因在布氏锥虫基因组中均以单拷贝形式存在。脉冲场凝胶分析表明,这些基因编码于不同的染色体上,因此不属于同一基因簇。这些亚基的蛋白质水平与稳态转录本水平之间的比较表明,表达调控主要发生在转录后。对前循环型和血流型mRNA稳定性的比较显示,这三种转录本在血流型中的半衰期要短得多。9号亚基在前循环型中转录本稳定性的差异大于α和β亚基,而蛋白质水平的差异相当。这些结果表明,9号亚基可能存在进一步的转录后调控。