Data for Figures 6 B, D, E, F, G
Description
Ras activity buffers extrinsic fluctuations that affect cell cycle dynamics B. Altering nutrition or pH during growth affects Ras network activity. Expression of rrgA and rigA was measured in G+ pH 6.8 and 7.5 and G- medium pH 6.8 by qPCR (3 replicates each). In stalky G- cells the expression of the low Ras activity reporter rrgA increases relative to high Ras activity reporter rigA expression. This trend is reversed in sporey alkaline pH. Error bars depict SEM. D. Increased stalk cell differentiation in rrgA- mutant cells when grown in G-. rrgA- mutant cells do not show a prestalk bias when grown in G+ medium, but the number of stalk cells is significantly increased compared to wild type cells when grown in G- medium. Error bars depict SEM. E. A rrgA and rigA dual promoter construct acts as a RasD network activity reporter. A plasmid containing the promoter of rrgA (RFP) and rigA(GFP) was transformed into wild type and gefE- cells. The majority of the cells (80.9% and 70%) express neither GFP nor RFP. In wild type, the majority of fluorescent cells predominantly express only GFP (15.1%). In the gefE- mutant, the majority of the fluorescent cells express RFP (20.6%). Only 3.3% and 2.2% express both markers, respectively. F and G. Cells that induce rrgA expression have a longer than average cell cycle. Cell cycle length was measured in cells transformed with the rrgA promoter-RFP reporter. Cells that induced the expression of rrgA have a slightly longer cell cycle, while the cell cycle of the mothers of rrgA expressing cells are significantly longer (one-way ANOVA, p=0.0019). Error bars depict SEM.