Fig. 5: T492 phosphorylation impacts heat-induced Ssa1-ribosome association and translational fidelity.

a mutation of T492 alters the interaction of Ssa1 with translational machinery. Data shown are taken from Fig. 3b. b ribosomal subunits displaying altered interaction upon T492 mutation. Structure based on PDB file 4V7R. c Polysome profiles of WT and T492A yeast under untreated and heat-shocked and recovery conditions. d Interaction of Ssa1, Ydj1 and Sis1 with polysome fractions in untreated, heat-shocked and recovery conditions. Each experiment involving western blotting was repeated three times with similar results e Translation was measured by incubating cells with the methionine analog homopropargylglycine (HPG) for the indicated time and then fluorescently labeling the cells with click chemistry. Control cells were incubated with the translation inhibitor cycloheximide (CHX). Statistical analysis was performed using two-way ANOVA. *p ≤ 0.05, **p ≤ 0.01. Data plotted is the mean fluorescence intensity of the population, normalized to WT (10 min) for each temperature. f Translational fidelity of WT and T492 cells was determined using a UUG/AUG dual luciferase assay as described in ref. ⁵¹. The data shown are the mean and standard deviation of three biological replicates. Statistical significance was calculated via a two-sided unpaired t test. **, P ≤ 0.01. Source data are provided as a Source Data file.