Mature tRNATrp levels are rescued upon met22 deletion in the trm10Δ strain. (A) Northern analysis of RNA derived from the indicated strains probed for mature tRNATrp, mature tRNAGly, and 5S rRNA, which were all detected using 5′ radiolabeled probes (Supplemental Table S2). Strains shown in lanes 1–8 are wild-type for MET22; whereas strains shown in lanes 9–16 are met22Δ strains, with lanes 13–16 also containing a CEN URA3 MET22 plasmid for complementation of met22Δ. A representative northern blot visualized at lower exposure is shown in Supplemental Figure S1C for comparison. (B) Quantification of relative mature tRNA levels from strains shown in A. Normalized RNA levels (to 5S rRNA) were calculated for each strain, and for lanes 1–8, lane 3 was set to 1 for determining relative tRNA levels, and for lanes 9–16, lane 15 was set to 1 for determining relative tRNA levels, in order to better visualize the positive and negative changes associated with each set of strains on the same axis. Triplicate data were plotted, with individual data points for tRNAGly(GCC) and tRNATrp shown. Two-tailed t-tests assuming equal variance were performed, and one-tailed P-value is represented by asterisks comparing tRNATrp levels in the trm10Δ and the trm10Δmet22Δ strain. P-values for comparisons not shown here are listed in Supplemental Table S1.
