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. Author manuscript; available in PMC: 2012 Dec 9.
Published in final edited form as: Mol Cell. 2011 Dec 9;44(5):745–758. doi: 10.1016/j.molcel.2011.09.019

Figure 5. Translational resurrection is restricted to a subset of genes for a limited time.

Figure 5

(A) Polysome profiles of cells subjected to 10 minutes of glucose starvation followed by 5 minutes of glucose repletion in the presence (+T) or absence of thiolutin to inhibit new transcription. Thiolutin treatment slightly reduced polysome recovery. Note the relative heights of the disome (open arrowhead) and polysome (filled arrowhead) peaks and the widths of the monosome peaks ± thio. Analytical polysome assays (A, E, F) were repeated 2–4 times. Representative traces are shown for each. (B) Ratio values shown by color scale from microarray analysis of non-polysomal (NP) and polysomal (P) mRNA from cells starved for 10 minutes (YP 10′ +thio) or starved and re-fed for 5 minutes (+D 5′ +thio). Polysomal mRNA from YPD cultures served as the reference sample for each array. Genes are organized according to T/T and P/T groups from Figure 2. Group 5 is shown at 50% vertical scale. Genes that showed low P/T ratios in starved cells were less depleted from the non-polysomal fraction than genes with high or neutral P/T ratios and showed greater mobilization into polysomes upon re-feeding. “Δpoly” values were derived from the ratio of ratios (YPD5_P versus YP10_P). (C) Graphs show quantification of the Δpoly values from B. Error bars in C and D indicate SEM. Asterisks (*) indicate Student’s t-test p-value < 0.0001. (D) RPGs as a class preferentially recovered in polysomes upon glucose re-addition. “All” = all genes from T/T 5–7. (E) Prolonged glucose starvation in the absence of new transcription leads to reduced polysome recovery upon re-feeding. Cells were starved for 45 (top row) or 60 (bottom row) minutes in the presence of thiolutin before re-feeding for 5 minutes. Polysome profiles of recovery (+D 5′) after only 10 minutes –glucose +thio are overlaid for comparison (red lines). (F) New transcription contributes substantially to polysome recovery after prolonged starvation. Polysome profiles of recovery (+D 5′) after 45′ or 60′ of starvation are shown in black. Polysome recovery after starvation for 45 or 60 minutes +thiolutin is shown (blue lines) for comparison. See also Figure S5.