Effect of fosmidomycin on metabolic and transcript profiles of the methylerythritol phosphate pathway in Plasmodium falciparum
Effect of fosmidomycin on metabolic and transcript profiles of the methylerythritol phosphate pathway in Plasmodium falciparum
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DOI：
10.1590/s0074-02762007000300019
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发表时间：
2007-06-01
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影响因子：
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通讯作者：
Katzin, Alejandro M
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文献类型：
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作者：
Cassera, María B;Merino, Emilio F;Katzin, Alejandro M
在恶性疟原虫中，异戊二烯基二磷酸和二甲基烯丙基二磷酸是类异戊二烯生物合成的中心中间体，其形成是通过甲基赤四醇磷酸（MEP）途径进行的。Fosmidomycin是MEP途径的第二种酶，1-脱氧-d -木醛糖-5-磷酸还原异构酶的特异性抑制剂。我们分析了在整个恶性疟原虫红细胞周期中，磷霉素对每种中间体水平的影响及其对类异戊二烯生物合成（如酚类和泛醌类）的代谢需求。通过实时聚合酶链反应定量MEP通路相关基因的稳态RNA水平，并与相关代谢物水平进行相关性分析。我们的研究结果表明，在红细胞周期中，MEP途径代谢产物的峰值先于转录物的最大丰度。fosmidomycin处理导致MEP通路的中间水平降低，以及泛醌和醇的生物合成。药物适度改变了MEP通路相关转录本，表明寄生虫在转录水平上没有强烈的反应。这是第一个比较fosmidomycin对恶性疟原虫代谢和转录谱影响的研究，恶性疟原虫只有MEP途径进行类异戊二烯生物合成。
In Plasmodium falciparum, the formation of isopentenyl diphosphate and dimethylallyl diphosphate, central intermediates in the biosynthesis of isoprenoids, occurs via the methylerythritol phosphate ( MEP) pathway. Fosmidomycin is a specific inhibitor of the second enzyme of the MEP pathway, 1-deoxy-D-xylulose-5-phosphate reductoisomerase. We analyzed the effect of fosmidomycin on the levels of each intermediate and its metabolic requirement for the isoprenoid biosynthesis, such as dolichols and ubiquinones, throughout the intraerythrocytic cycle of P. falciparum. The steady-state RNA levels of the MEP pathway-associated genes were quantified by real-time polymerase chain reaction and correlated with the related metabolite levels. Our results indicate that MEP pathway metabolite peak precede maximum transcript abundance during the intraerythrocytic cycle. Fosmidomycin-treatment resulted in a decrease of the intermediate levels in the MEP pathway as well as in ubiquinone and dolichol biosynthesis. The MEP pathway associated transcripts were modestly altered by the drug, indicating that the parasite is not strongly responsive at the transcriptional level. This is the first study that compares the effect of fosmidomycin on the metabolic and transcript profiles in P. falciparum, which has only the MEP pathway for isoprenoid biosynthesis.