Čáp Michal, Váchová Libuše, Palková Zdena
a Department of Genetics and Microbiology ; Faculty of Science; Charles University in Prague ; Prague , Czech Republic.
b Institute of Microbiology of the Academy of Sciences of the Czech Republic ; Prague , Czech Republic.
Cell Cycle. 2015;14(21):3488-97. doi: 10.1080/15384101.2015.1093706.
Colonies of Saccharomyces cerevisiae laboratory strains pass through specific developmental phases when growing on solid respiratory medium. During entry into the so-called alkali phase, in which ammonia signaling is initiated, 2 prominent cell types are formed within the colonies: U cells in upper colony regions, which have a longevity phenotype and activate the expression of a large number of metabolic genes, and L cells in lower regions, which die more quickly and exhibit a starvation phenotype. Here, we performed a detailed analysis of the activities of enzymes of central carbon metabolism in lysates of both cell types and determined several fermentation end products, showing that previously reported expression differences are reflected in the different enzymatic capabilities of each cell type. Hence, U cells, despite being grown on respiratory medium, behave as fermenting cells, whereas L cells rely on respiratory metabolism and possess active gluconeogenesis. Using a spectrum of different inhibitors, we showed that glycolysis is essential for the formation, and particularly, the survival of U cells. We also showed that β-1,3-glucans that are released from the cell walls of L cells are the most likely source of carbohydrates for U cells.
酿酒酵母实验室菌株的菌落在固体呼吸培养基上生长时会经历特定的发育阶段。在进入所谓的碱性阶段(在此阶段启动氨信号传导)期间,菌落在内部形成两种突出的细胞类型:菌落上部区域的U细胞,具有长寿表型并激活大量代谢基因的表达;以及下部区域的L细胞,其死亡更快并表现出饥饿表型。在这里,我们对这两种细胞类型裂解物中的中心碳代谢酶活性进行了详细分析,并测定了几种发酵终产物,结果表明先前报道的表达差异反映在每种细胞类型的不同酶促能力上。因此,U细胞尽管生长在呼吸培养基上,却表现为发酵细胞,而L细胞则依赖呼吸代谢并具有活跃的糖异生作用。使用一系列不同的抑制剂,我们表明糖酵解对于U细胞的形成,特别是存活至关重要。我们还表明,从L细胞细胞壁释放的β-1,3-葡聚糖最有可能是U细胞的碳水化合物来源。