Impact of Tda1 on gene expression during glycerol cultivation in Saccharomyces cerevisiae

The energy metabolism pathways are significantly influenced by the available carbon sources. In Saccharomyces cerevisiae, energy is primarily produced through aerobic respiration during glycerol cultivation, a process believed to depend on the yeast AMPK (AMP-activated protein kinase) homolog, Snf1. It has been reported that Snf1 increases the expression of respiratory genes by phosphorylating transcriptional activators in environments where glucose is unavailable. We discovered that Tda1, activated by Snf1, phosphorylates Hxk2. Hxk2 has been reported to function as a transcriptional repressor. Therefore, we analyzed how Tda1 affects the expression of respiratory genes by RNA-seq analysis of glycerol-cultured cells. Overall design: RNA-seq profiling of BY4741 WT, tda1?, snf1?, and Hxk2-S15A mutant cells under glycerol culture conditions.

能量代谢通路显著受可利用碳源的调控。在酿酒酵母(Saccharomyces cerevisiae)中，细胞在甘油培养条件下主要通过有氧呼吸产生能量，这一过程被认为依赖于酵母AMP活化蛋白激酶(AMP-activated protein kinase, AMPK)的同源蛋白Snf1。已有研究表明，在葡萄糖匮乏的环境中，Snf1可通过磷酸化转录激活因子提升呼吸相关基因的表达水平。本研究发现，经Snf1激活的Tda1可磷酸化Hxk2；已有研究显示Hxk2可作为转录阻遏蛋白发挥功能。据此，本研究通过对甘油培养的细胞开展RNA测序（RNA-seq）分析，探究了Tda1对呼吸相关基因表达的调控作用。整体实验设计：在甘油培养条件下，对BY4741野生型（WT）、tda1缺失株、snf1缺失株及Hxk2-S15A突变体细胞进行RNA-seq转录组分析。