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Targeted proteome analysis of single-gene deletion strains of Saccharomyces cerevisiae lacking enzymes in the central carbon metabolism

机译:在中央碳代谢中缺乏酶的酿酒酵母单基因缺失菌株的靶向蛋白质组分析

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摘要

Central carbon metabolism is controlled by modulating the protein abundance profiles of enzymes that maintain the essential systems in living organisms. In this study, metabolic adaptation mechanisms in the model organism Saccharomyces cerevisiae were investigated by direct determination of enzyme abundance levels in 30 wild type and mutant strains. We performed a targeted proteome analysis using S. cerevisiae strains that lack genes encoding the enzymes responsible for central carbon metabolism. Our analysis revealed that at least 30% of the observed variations in enzyme abundance levels could be explained by global regulatory mechanisms. A enzyme-enzyme co-abundance analysis revealed that the abundances of enzyme proteins involved in the trehalose metabolism and glycolysis changed in a coordinated manner under the control of the transcription factors for global regulation. The remaining variations were derived from local mechanisms such as a mutant-specific increase in the abundances of remote enzymes. The proteome data also suggested that, although the functional compensation of the deficient enzyme was attained by using more resources for protein biosynthesis, available resources for the biosynthesis of the enzymes responsible for central metabolism were not abundant in S. cerevisiae cells. These results showed that global and local regulation of enzyme abundance levels shape central carbon metabolism in S. cerevisiae by using a limited resource for protein biosynthesis.
机译:中央碳代谢通过调节维持生命有机体必不可少的系统的酶的蛋白质丰度分布来控制。在这项研究中,通过直接测定30种野生型和突变菌株中酶的丰度水平,研究了模型生物酿酒酵母中的代谢适应机制。我们使用酿酒酵母菌株进行了靶向蛋白质组分析,该菌株缺乏编码负责中央碳代谢的酶的基因。我们的分析表明,至少30%的酶丰度水平变化可以通过全球调控机制来解释。酶-酶共丰度分析表明,参与海藻糖代谢和糖酵解的酶蛋白质的丰度在转录因子的控制下以整体方式协调地变化。其余的变异来自局部机制,例如远程酶丰度的突变体特异性增加。蛋白质组数据还表明,尽管通过使用更多的蛋白质生物合成资源来获得缺陷酶的功能性补偿,但是在酿酒酵母细胞中,用于负责中央代谢的酶的生物合成的可用资源并不丰富。这些结果表明,通过使用有限的蛋白质生物合成资源,酶的丰度水平的整体和局部调节会影响酿酒酵母的中央碳代谢。

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