Fig. 2.

Comparison of effects of Sui⁻ and Ssu⁻ mutations in eIF1, eIF2β, and eIF1A on eIF1 protein and SUI1 mRNA levels. (A) Northern analysis of SUI1 mRNA in selected mutants. Lanes 1 to 6 and lanes 12 to 17 show results for strains described in Fig. 1C and D and in Fig. 5A, B, and F containing the indicated SUI1⁺ alleles (lanes 1 to 6) or SUI1-opt alleles (lanes 12 to 17) cultured as in Fig. 1C. Lanes 7 to 11 and lanes 18 to 22 show the results for strains described in Fig. 6A and C containing SUI1⁺ (lanes 7 to 11) or SUI1-opt (lanes 18 to 22) cultured as described in Fig. 6A. Total RNA was subjected to Northern analysis of SUI1 and PYK1 mRNAs, the hybridization signals were quantified with a Phosphorimager, and ratios of SUI1 to PYK1 mRNA were calculated and normalized to the ratio obtained for the corresponding strain, harboring WT SUI1, SUI3, and TIF11. The resulting “Norm. SUI1/PYK1” values are listed below the blot and also in columns 3 and 7 of panel B. The WT SUI1 reference strain is shown in lane 1 for the mutants examined in lanes 1 to 6 and lanes 12 to 17. The WT SUI1 SUI3 reference strain is shown in lane 7 for the mutants examined in lanes 7 and 8 and lanes 18 and 19. The WT SUI1 TIF11 reference strain is shown in lane 9 for the mutants examined in lanes 9 to 11 and lanes 20 to 22. (B) Comparison of eIF1 protein and SUI1 mRNA levels. For the strains analyzed in panel A, the appropriate “Rel. eIF1/Gcd6” values taken from Fig. 1C and D, from Fig. 5A, B, and F, and from Fig. 6A and C are listed in columns 2 and 6 (eIF1/Gcd6 Protein) for SUI1 and SUI1-opt alleles, respectively; the “Norm. SUI1/PYK1” values from panel A are listed in columns 3 and 7 (SUI1/PYK1 mRNA); the ratios of values in columns 2 and 3 are listed in column 4 (Protein/mRNA); the ratios of values in columns 6 and 7 are listed in column 8 (Protein/mRNA); and the values in column 8 normalized to the cognate WT ratios in rows 12, 18, or 20 are listed in column 9 (Norm. Protein/mRNA).