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. 2019 Jun 3;2(3):e201800261. doi: 10.26508/lsa.201800261

Figure 3. In the context of stress, methylation of Rpc31 is required for robust repression of pre-tRNA biogenesis.

Figure 3.

(A, B) RNA hybridization was carried out for four pre-tRNAs in WT versus hmt1Δ cells (A) or WT versus Rpc31R5,9A cells (B) grown in YPD before and after treatment with CPZ. Ratios of signal intensities for each pre-tRNA were individually normalized against three internal controls: U4, U3, and U5. The normalized signals were plotted on a bar graph to compare against signal obtained from the untreated WT cells, which is set to 100%. Error bars represent the SEM of three biological replicates (n = 3). P-value as calculated by t test: *<0.05, **<0.01, and ***<0.001. (C) Fold decrease in expression of four candidate pre-tRNAs in WT, hmt1Δ, or Rpc31R5,9A cells after treatment with CPZ, as assessed by hybridization of probes to intronic regions. Signal was normalized to levels of the U4 snRNA. Analysis of variance (ANOVA) revealed significant variation among the strains(P-value = 1.4 × 10−6). Post hoc Tukey’s Honest significant differences method revealed a significant difference between WT and hmt1Δ (after adjustment for the multiple comparisons, the adjusted P-value is 0.0014), WT and Rpc31R5,9A (adjusted P-value is 2.8 × 10−6). n = at least three per pre-tRNA.