Figure 4.

Transcriptome analysis of strains expressing mistranslating tRNA variants. (A) PCA of centered log ratio normalized reads from BY4742 expressing either ${\text{tRNA}}_{\mathrm{G}3\text{:}\mathrm{U}70}^{\text{Pro}}$ (Pro→Ala), ${\text{tRNA}}_{\text{UCU},\mathrm{G}26\mathrm{A}}^{\text{Ser}}$ (Arg→Ser) or an empty vector (WT). Each point represents one biological replicate (n = 3). (B) Heatmap of hierarchical clustered differentially expressed genes (P ≤ 0.05) in response to mistranslation. Fold-change for each gene is the average from three biological replicates. Upregulated genes are colored yellow while downregulated genes are colored blue. Significantly enriched GO terms within each cluster are annotated. (C) Venn diagram of upregulated and downregulated genes (P ≤ 0.05) in yeast strain BY4742 transformed with a centromeric plasmid expressing ${\text{tRNA}}_{\mathrm{G}3\text{:}\mathrm{U}70}^{\text{Pro}}$ (Pro→Ala) or ${\text{tRNA}}_{\text{UCU},\mathrm{G}26\mathrm{A}}^{\text{Ser}}$ (Arg→Ser) as compared to BY4742 transformed with empty vector. (D) Volcano plot highlighting in yellow differentially expressed genes with greater than twofold changes in ${\text{tRNA}}_{\mathrm{G}3\text{:}\mathrm{U}70}^{\text{Pro}}$ relative to the control strain. Points with gene name labels have the largest fold-change relative to control. (E) Volcano plot highlighting in blue differentially expressed genes with greater than twofold changes in ${\text{tRNA}}_{\text{UCU},\mathrm{G}26\mathrm{A}}^{\text{Ser}}$ as in D.