Wu Jian, Zhang Nianshu, Hayes Andrew, Panoutsopoulou Kalliope, Oliver Stephen G
School of Biological Sciences, University of Manchester, 2.205 Stopford Building, Oxford Road, Manchester M13 9PT, United Kingdom.
Proc Natl Acad Sci U S A. 2004 Mar 2;101(9):3148-53. doi: 10.1073/pnas.0308321100. Epub 2004 Feb 18.
Global gene expression in yeast was examined in five different nutrient-limited steady states and in their corresponding starvation-induced stationary phases. The use of chemostats, with their ability to generate defined and reproducible physiological conditions, permitted the exclusion of the confounding variables that frequently complicate transcriptome analyses. This approach allowed us to dissect out effects on gene expression that are specific to particular physiological states. Thus, we discovered that a large number of ORFs involved in protein synthesis were activated under ammonium limitation, whereas the expression of ORFs concerned with energy and metabolism was enhanced by carbon limitation. Elevated transcription of genes in high-affinity glucose uptake, the trichloroacetic acid cycle, and oxidative phosphorylation were observed in glucose-limiting, but not glucose-abundant, conditions. In contrast, genes involved in gluconeogenesis and, interestingly, genes subject to nitrogen catabolite repression increased their transcription when ethanol was the carbon source, even though ammonium was in excess. This result suggests that up-regulation of genes sensitive to nitrogen catabolite repression may contribute anapleurotic intermediates in ethanol-grown cells. The different starvation conditions produced two general types of transcription profiles, with carbon-starved cells transcribing far fewer genes than cells starved for any of the other macronutrients. Nonetheless, each starvation condition induced its own peculiar set of genes, and only 17 genes were induced >5-fold by all five starvations. In all cases, analysis of the upstream sequences of clusters of coregulated genes identified motifs that may be recognized by transcription factors specific for controlling gene expression in each of the physiological conditions examined.
在五种不同的营养限制稳态及其相应的饥饿诱导静止期对酵母中的全局基因表达进行了检测。恒化器能够产生明确且可重复的生理条件,利用它可以排除那些经常使转录组分析复杂化的混杂变量。这种方法使我们能够剖析出特定生理状态对基因表达的影响。因此,我们发现大量参与蛋白质合成的开放阅读框(ORF)在铵限制下被激活,而与能量和代谢相关的开放阅读框的表达在碳限制下增强。在葡萄糖限制而非葡萄糖丰富的条件下,观察到高亲和力葡萄糖摄取、三羧酸循环和氧化磷酸化相关基因的转录升高。相反,当乙醇作为碳源时,即使铵过量,参与糖异生的基因以及有趣的是受氮分解代谢物阻遏的基因的转录会增加。这一结果表明,对氮分解代谢物阻遏敏感的基因上调可能为乙醇生长的细胞提供回补中间产物。不同的饥饿条件产生了两种一般类型的转录谱型,碳饥饿的细胞转录的基因比其他任何常量营养素饥饿的细胞少得多。尽管如此,每种饥饿条件都诱导了其特有的一组基因,并且只有17个基因在所有五种饥饿条件下均被诱导超过5倍。在所有情况下,对共调控基因簇上游序列的分析都鉴定出了可能被在所检测的每种生理条件下控制基因表达的特异性转录因子识别的基序。