Meijer M M, Boonstra J, Verkleij A J, Verrips C T
Utrecht University, Department of Molecular Cell Biology/Institute for Biomembranes, Padualaan 8, 3584 CH Utrecht, The Netherlands.
J Biol Chem. 1998 Sep 11;273(37):24102-7. doi: 10.1074/jbc.273.37.24102.
Glucose plays an important regulatory role in the yeast Saccharomyces cerevisiae, which is mostly reflected at the transcriptional level by glucose repression. The signal that initiates glucose repression is unknown, but data indicate that it is located at or above the level of glucose 6-phosphate, suggesting the involvement of either the intracellular or extracellular glucose concentration or the glucose flux in triggering glucose repression. We have investigated the role of the glucose flux and the extracellular glucose concentration in glucose repression by growing the cells in continuous culture under nitrogen limitation. By a step-wise increase in the glucose feed concentration, the glucose flux and extracellular glucose concentrations were modulated in an accurate way. Furthermore, the glucose flux and glucose concentrations were modulated independently of each other by increasing the dilution rate or by the use of fructose as a substrate. Using these approaches we demonstrate that glucose repression is related to the extracellular (or intracellular) glucose concentration rather than the glucose flux. At external glucose concentrations lower than 14 mM, glucose repression of SUC2 gene transcription was not triggered, whereas glucose repression of this gene was activated when the glucose concentration exceeded 18 mM. A comparable effect was observed for the glucose-repressible carbon source fructose.
葡萄糖在酿酒酵母中发挥着重要的调节作用,这主要通过葡萄糖阻遏在转录水平上得以体现。启动葡萄糖阻遏的信号尚不清楚,但数据表明该信号位于6-磷酸葡萄糖水平或之上,这表明细胞内或细胞外葡萄糖浓度或葡萄糖通量参与触发葡萄糖阻遏。我们通过在氮限制条件下进行连续培养来研究葡萄糖通量和细胞外葡萄糖浓度在葡萄糖阻遏中的作用。通过逐步提高葡萄糖进料浓度,可精确调节葡萄糖通量和细胞外葡萄糖浓度。此外,通过提高稀释率或使用果糖作为底物,可使葡萄糖通量和葡萄糖浓度相互独立地进行调节。利用这些方法,我们证明葡萄糖阻遏与细胞外(或细胞内)葡萄糖浓度相关,而非与葡萄糖通量相关。当外部葡萄糖浓度低于14 mM时,未触发SUC2基因转录的葡萄糖阻遏,而当葡萄糖浓度超过18 mM时,该基因的葡萄糖阻遏被激活。对于可被葡萄糖阻遏的碳源果糖,也观察到了类似的效应。