Boer Viktor M, de Winde Johannes H, Pronk Jack T, Piper Matthew D W
Kluyver Laboratory of Biotechnology, Technical University of Delft, Julianalaan 67, 2628BC Delft, The Netherlands.
J Biol Chem. 2003 Jan 31;278(5):3265-74. doi: 10.1074/jbc.M209759200. Epub 2002 Oct 31.
Profiles of genome-wide transcriptional events for a given environmental condition can be of importance in the diagnosis of poorly defined environments. To identify clusters of genes constituting such diagnostic profiles, we characterized the specific transcriptional responses of Saccharomyces cerevisiae to growth limitation by carbon, nitrogen, phosphorus, or sulfur. Microarray experiments were performed using cells growing in steady-state conditions in chemostat cultures at the same dilution rate. This enabled us to study the effects of one particular limitation while other growth parameters (pH, temperature, dissolved oxygen tension) remained constant. Furthermore, the composition of the media fed to the cultures was altered so that the concentrations of excess nutrients were comparable between experimental conditions. In total, 1881 transcripts (31% of the annotated genome) were significantly changed between at least two growth conditions. Of those, 484 were significantly higher or lower in one limitation only. The functional annotations of these genes indicated cellular metabolism was altered to meet the growth requirements for nutrient-limited growth. Furthermore, we identified responses for several active transcription factors with a role in nutrient assimilation. Finally, 51 genes were identified that showed 10-fold higher or lower expression in a single condition only. The transcription of these genes can be used as indicators for the characterization of nutrient-limited growth conditions and provide information for metabolic engineering strategies.
对于给定环境条件下的全基因组转录事件概况,在诊断定义不明确的环境中可能具有重要意义。为了识别构成此类诊断概况的基因簇,我们对酿酒酵母在碳、氮、磷或硫限制下生长的特定转录反应进行了表征。使用在恒化器培养中以相同稀释率在稳态条件下生长的细胞进行微阵列实验。这使我们能够研究一种特定限制的影响,而其他生长参数(pH、温度、溶解氧张力)保持不变。此外,改变供给培养物的培养基组成,以便在实验条件之间过量营养物的浓度具有可比性。总共,1881个转录本(注释基因组的31%)在至少两种生长条件之间有显著变化。其中,484个仅在一种限制条件下显著升高或降低。这些基因的功能注释表明细胞代谢发生改变以满足营养限制生长的生长需求。此外,我们确定了几种在营养同化中起作用的活性转录因子的反应。最后,鉴定出51个基因,它们仅在单一条件下表达升高或降低10倍。这些基因的转录可以用作表征营养限制生长条件的指标,并为代谢工程策略提供信息。