Regulation of GCN4 translation by uORFs. (A) The wild-type and gcn2 mutant strains containing a wild-type GCN4-Luc construct were treated with 15 mM 3AT for 3 h. The uORFs in GCN4 are indicated as boxes in the schematic. (B) The wild-type and gcn2 mutant strains containing a GCN4-Luc construct where uORF1–3 were mutated were treated with 15 mM 3AT for 1 h. Mutations in the uORFs of GCN4 are indicated as crosses in the schematic. (C) Quantitative RT-PCR analysis of GCN4-Luc mRNA levels relative to GAPDH mRNA levels. (D) Quantitative RT-PCR analysis of GCN4 mRNA levels relative to GAPDH mRNA levels. White-colored bars denote values obtained from nonstressed cells; black-colored bars, values obtained from cells treated with 15 mM 3AT for 3 h.
