Skip to main content
. 2021 Aug 5;49(15):8743–8756. doi: 10.1093/nar/gkab652

Figure 3.

Figure 3.

Growth analysis of the selected eIF4G mutants with or without overexpression of eIF4A or eIF4E. (A) Schematic representation of the eIF4G1 (adopted from (34)) showing the positions of introduced point mutations and changes in amino acid residues analyzed in this study. (B) Western blot analysis of WCEs from the strains described in panel C grown at 30°C, using anti-His, anti-eIF4A, and anti-eIF4E antibodies. (C) The eIF4G1-1, eIF4G1-8 and eIF4G1-459 mutant strains impart the Ts– and the latter also Slg– phenotypes. Strain YMP47 (GCN2 tif4631Δ, tif4632Δ) was transformed with individual YEplac181-based plasmids carrying the eIF4G1 alleles and the resident YEp-eIF4G1-URA plasmid was evicted on 5-FOA. The resulting strains, eIF4G1 (YMP53), eIF4G1-1 (YMP55), -8 (YMP56), -8* (YMP91) and -459 (YMP57), together with the isogenic H199 strain (GCN2 TIF4631, TIF4631) were then spotted in five serial 10-fold dilutions on SD plates and incubated at 30°C or 34°C for 3 days. (D) Growth defects of individual eIF4G mutants are suppressible by either high copy expression of eIF4E or eIF4A. The strains described in panel C were transformed individually with YEplac112-based plasmids carrying either eIF4A or eIF4E alleles, or an empty plasmid, and the resulting transformants were spotted in five serial 10-fold dilutions on SD plates and incubated at 30 or 34°C for 3 days.