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. 2003 Apr 1;17(7):859–872. doi: 10.1101/gad.1069003

Figure 5.

Figure 5

(A,B) The tap42-11 mutation alters the effects of rapamycin on phosphorylation of Ser 577 in GCN2 and Ser 51 in eIF2α at 30°C. Experiments analogous to those described in Figure 4A and B were carried out identically except that the strains were grown to early logarithmic phase at 25°C in SC medium lacking uracil or SC complete and transferred to the same medium prewarmed to 30°C for 30 min, after which they were treated with 200 ng/mL rapamycin for 20 min. (C,D) Effects of inactivating TAP42-regulated phosphatases on dephosphorylation of S577 and eIF2α phosphorylation. Transformants of strains BY4741 (WT), #3744 (sit4Δ), and CY1090 (pph21Δ pph22Δ) containing GCN2-FL on pDH101, were grown on SC medium lacking uracil to early logarithmic phase at 30°C and treated with 200 ng/mL rapamycin for 20 min, or were left untreated. Phosphorylation of Ser 577 in FL-GCN2 (C) or Ser 51 in eIF2α (D) was detected and quantified as described in Figure 1. Strain BY4741 transformed with pCB149 containing GCN2-S577A-FL (C) or gcn2Δ strain ESY6053 transformed with empty vector (D) was analyzed in parallel (lane 1).