Figure 6: Loss of Efm methyltransferases and mutation of four lysine residues to arginine residues in EF1A results in differential responses to translational inhibitors.

Representative image of yeast cells grown in YPD and then serially diluted onto agar plates as described in the Figure 3 legend but supplemented with either puromycin, cycloheximide, tunicamycin and anisomycin. In puromycin, 8 out of 10 replicates for efm14567Δ, and 10 out of 12 replicates for efm1456Δ strain had smaller colonies compared to wildtype colonies. Colonies for the TEF1 K(30,79,316,390)R strain always had similar sized colonies compared to wildtype colonies in 12 replicates. Growth on cycloheximide displayed no difference in colony size compared to wildtype colonies for the efm14567Δ mutant (four replicate experiments), efm1456Δ mutant (eight replicate experiments), and the TEF1 K(30,79,316,390)R mutant (eight replicate experiments). Tunicamycin colony sizes were always smaller than wildtype for efm1456Δ (six replicate experiments) and efm14567Δ (two replicated experiments) but remained unchanged for TEF1 K(30,79,316,390)R mutant. (six replicate experiments). On anisomycin plates, there were smaller colonies 4 out of 6 replicates for efm14567Δ and 5 out of 6 replicates for the efm1456Δ mutant compared to wildtype colonies. The TEF1 K(30,79,316,390)R strain had similar sized colonies compared to wildtype colonies with the exception of 2 out of 6 replicates where the colony sizes were bigger.