Boer Viktor M, Tai Siew Leng, Vuralhan Zeynep, Arifin Yalun, Walsh Michael C, Piper Matthew D W, de Winde Johannes H, Pronk Jack T, Daran Jean-Marc
Department of Biotechnology, Delft University of Technology, Julianalaan, Delft, The Netherlands.
FEMS Yeast Res. 2007 Jun;7(4):604-20. doi: 10.1111/j.1567-1364.2007.00220.x. Epub 2007 Apr 10.
Aerobic, glucose-limited chemostat cultures of Saccharomyces cerevisiae grown with six different nitrogen sources were subjected to transcriptome analysis. The use of chemostats enabled an analysis of nitrogen-source-dependent transcriptional regulation at a fixed specific growth rate. A selection of preferred (ammonium and asparagine) and nonpreferred (leucine, phenylalanine, methionine and proline) nitrogen sources was investigated. For each nitrogen source, distinct sets of genes were induced or repressed relative to the other five nitrogen sources. In total, 131 such 'signature transcripts' were identified in this study. In addition to signature transcripts, genes were identified that showed a transcriptional coresponse to two or more of the six nitrogen sources. For example, 33 genes were transcriptionally upregulated in leucine-grown, phenylalanine-grown and methionine-grown cultures; this was partly attributed to the involvement of common enzymes in the dissimilation of these amino acids. In addition to specific transcriptional responses elicited by individual nitrogen sources, their impact on global regulatory mechanisms such as nitrogen catabolite repression (NCR) were monitored. NCR-sensitive gene expression in the chemostat cultures showed that ammonium and asparagine were 'rich' nitrogen sources. By this criterion, leucine, proline and methionine were 'poor' nitrogen sources, and phenylalanine showed an 'intermediate' NCR response.
对在六种不同氮源下进行有氧、葡萄糖受限恒化培养的酿酒酵母进行了转录组分析。使用恒化器能够在固定的比生长速率下分析氮源依赖性转录调控。研究了一系列首选(铵和天冬酰胺)和非首选(亮氨酸、苯丙氨酸、蛋氨酸和脯氨酸)氮源。对于每种氮源,相对于其他五种氮源,诱导或抑制了不同的基因集。在本研究中总共鉴定出131个这样的“特征转录本”。除了特征转录本外,还鉴定出了对六种氮源中的两种或更多种表现出转录协同反应的基因。例如,在亮氨酸培养、苯丙氨酸培养和蛋氨酸培养的培养物中有33个基因转录上调;这部分归因于这些氨基酸异化过程中常见酶的参与。除了单个氮源引发的特定转录反应外,还监测了它们对诸如氮分解代谢物阻遏(NCR)等全局调控机制的影响。恒化器培养物中对NCR敏感的基因表达表明,铵和天冬酰胺是“丰富”的氮源。根据这一标准,亮氨酸、脯氨酸和蛋氨酸是“贫”氮源,而苯丙氨酸表现出“中等”的NCR反应。