TORC1 and PKA dependent gene regulation. TORC1 and PKA dependent gene regulation

To determine how TORC1 and PKA cooperate to regulate cell growth, we performed temporal analysis of gene expression in yeast switched from a non-fermentable substrate, to glucose, in the presence and absence of TORC1 and PKA inhibitors. Quantitative analysis of these data reveals that PKA drives the expression of key cell growth genes during transitions into, and out of, the rapid growth state in glucose, while TORC1 is important for the steady-state expression of the same genes. Overall design: The study includes 21 experiments that measure the impact that TORC1 and PKA kinases have on gene expression as cells transition from growth in glycerol, to growth in glucose. All experiments were carried out using the cell line (ACY142), or a derivative of ACY142 (missing Dot6 and Tod6), that is carrying mutations in TPK1, 2 and 3 that render them sensitive to the Shokat inhibitor 1-NM-PP1.

为解析雷帕霉素靶蛋白复合物1（TORC1）与蛋白激酶A（PKA）协同调控细胞生长的分子机制，我们对以非发酵底物培养后切换至葡萄糖培养基的酵母开展了时序基因表达分析，实验分别设置了TORC1与PKA抑制剂存在、缺失两组条件。对上述数据的定量分析表明，在细胞进入及脱离葡萄糖快速生长状态的过程中，PKA驱动关键细胞生长基因的表达，而TORC1则对这些基因的稳态表达至关重要。整体设计：本研究共包含21组实验，旨在探究TORC1与PKA激酶对细胞从甘油生长环境切换至葡萄糖生长环境过程中基因表达的影响。所有实验均采用ACY142细胞系或其缺失Dot6与Tod6的衍生株，该细胞系携带TPK1、TPK2及TPK3突变，使其对Shokat抑制剂1-NM-PP1敏感。