Eukaryotic translation initiation factor eIF5 promotes the accuracy of start codon recognition by regulating Pi release and conformational transitions of the preinitiation complex.

Nucleic Acids Res. 2014, 42, 9623–40. doi: 10.1093/nar/gku653

The authors wish to make the following corrections to their article. The results and conclusion of the article are not affected and remain valid.

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In Figure 2A, row 4 (labelled G31R,M18V), the double mutant does not grow on -His medium in the presence of TIF5. Thus, it confers a dominant Ssu- phenotype.

Figure 2.

Genetic characterization of intragenic suppressors of TIF5 allele SUI5/G31R. (A) Derivatives of his4-301 tif5Δ strain ASY100 harboring a TIF5 plasmid and the indicated TIF5-FL alleles on LEU2 plasmids (hc WT on pAS5-132, sc WT on pAS5-101, G31R on pAS5-111, G31R,M18V on pAS5-115, G31R,K33E on pAS5-117, G31R,G62S on pAS5-112, G31R,L61A on pAS5-116 and hc G31R,T34N on pAS5-136) were replica-plated to SC-LU supplemented with either 0.3 mM histidine (-LU) or 0.0003 mM histidine (-LUH), or to SC-L supplemented with 5.2 mM 5-FOA (5-FOA). Cells were incubated for 3d (-LU) or 6d (-LUH and 5-FOA) at 30°C. (B) 10-fold serial dilutions of strains described in (A) were spotted on SC-LU and incubated for 3d at 30°C. (C) Derivatives of tif5Δ strain ASY137 harboring a TIF5 plasmid and the indicated TIF5 alleles on LEU2 plasmids were transformed with HIS4-lacZ reporter plasmids with AUG (p367) or UUG (p391) start codons. Cells were cultured in SC lacking leucine, tryptophan and uracil at 30°C and β-galactosidase activities were measured in whole cell extracts (WCEs). Ratios of β-galactosidase expressed from the UUG to AUG reporter were calculated from three independent transformants and mean ratios and and S.E.M.s (error bars) were plotted. (D) Slg− and His+/Sui− phenotypes of the his4-301 tif5Δ strains harboring TIF5 alleles described in (A), and isogenic strains containing M18V on pAS5-106, K33E on pAS5-108, G62S on pAS5-103, L61A on pAS5-107 or hc T34N on pAS5-135, were determined by spotting serial 10-fold dilutions on SC-LU supplemented with 0.3 mM His (+His) or 0.0003 mM His (−His) and incubated for 3d (−His) or 6d (+His) at 30°C. (E) Derivatives of his4-301 sui1Δ strain ASY250 with the chromosomal TIF5 gene under the GAL1 promoter (P) harboring SUI1+ on a TRP1 plasmid and the indicated TIF5 alleles on LEU2 plasmids were transformed with the AUG or UUG HIS4-lacZ reporter plasmids. Transformants were cultured in synthetic minimal medium with 2% galactose as carbon source and supplemented with 0.3 mM histidine (SGal+H) and then shifted to synthetic minimal medium with 2% glucose (SD+H) for 16 h. UUG:AUG initiation ratios for the HIS4-lacZ reporters were determined as in (C). (F) HIS4-lacZ UUG:AUG initiation ratios were measured as in (C) for strains described in (D) harboring the indicated plasmid-borne TIF5 alleles. For panels A and D, images have been cropped from results obtained from different plates examined in parallel in the same experiments.