Nishizawa Masafumi, Katou Yuki, Shirahige Katsuhiko, Toh-e Akio
Department of Microbiology and Immunology, Keio University School of Medicine, Shinjuku, Tokyo, Japan.
Yeast. 2004 Aug;21(11):903-18. doi: 10.1002/yea.1138.
The budding yeast Saccharomyces cerevisiae changes its gene expression profile when environmental nutritional conditions are changed. Protein kinases including cyclic AMP-dependent kinase, Snf1 and Tor kinases play important roles in this process. Pho85 kinase, a member of the yeast cyclin-dependent kinase family, is involved in the regulation of phosphate metabolism and reserve carbohydrates, and thus is implicated to function as a nutrient-sensing kinase. Upon depletion of glucose in the medium, yeast cells undergo a diauxic shift, accompanied by a carbon metabolic pathway shift, stimulation of mitochondrial function and downregulation of ribosome biogenesis and protein synthesis. We analysed the effect of a pho85Delta mutation on the expression profiles of the genes in this process to investigate whether Pho85 kinase participates in the yeast diauxy. We found that, in the absence of PHO85, a majority of mitochondrial genes were not properly induced, that proteasome-related and chaperonin genes were more repressed, and that, when glucose was still present in the medium, a certain class of genes involved in ribosome biogenesis (ribosomal protein and rRNA processing genes) was repressed, whereas those involved in gluconeogenesis and the glyoxylate cycle were induced. We also found that PHO85 is required for proper expression of several metal sensor genes and their regulatory genes. These results suggest that Pho85 is required for proper onset of changes in expression profiles of genes responsible for the diauxic shift.
当环境营养条件发生变化时,出芽酵母酿酒酵母会改变其基因表达谱。包括环磷酸腺苷依赖性激酶、Snf1和Tor激酶在内的蛋白激酶在这一过程中发挥着重要作用。Pho85激酶是酵母细胞周期蛋白依赖性激酶家族的成员之一,参与磷酸盐代谢和储备碳水化合物的调节,因此被认为具有营养感应激酶的功能。当培养基中的葡萄糖耗尽时,酵母细胞会经历二次生长转变,伴随着碳代谢途径的转变、线粒体功能的刺激以及核糖体生物合成和蛋白质合成的下调。我们分析了pho85Δ突变对这一过程中基因表达谱的影响,以研究Pho85激酶是否参与酵母的二次生长。我们发现,在缺乏PHO85的情况下,大多数线粒体基因不能被正常诱导,蛋白酶体相关基因和伴侣蛋白基因受到更强的抑制,并且当培养基中仍存在葡萄糖时,一类参与核糖体生物合成的基因(核糖体蛋白和rRNA加工基因)受到抑制,而参与糖异生和乙醛酸循环的基因则被诱导。我们还发现,PHO85是几个金属传感器基因及其调控基因正常表达所必需的。这些结果表明,Pho85是二次生长转变相关基因表达谱变化正常启动所必需的。