Levels of tRNATrp decrease upon deletion of trm10 and are almost entirely depleted upon growth of this strain in 5FU. (A) Northern analysis of RNA derived from the indicated strains grown with and without 5FU. Mature tRNATrp, mature tRNAGly(GCC), and 5S rRNA levels were all detected using 5′ radiolabeled probes (Supplemental Table S2). A representative northern blot using chemiluminescent detection is shown in Supplemental Figure S1B that displays the same RNAs at a lower exposure for comparison. (B) Quantification of relative tRNA levels from strains shown in A. Relative tRNA levels were calculated by comparing the observed intensity for each tRNA to the normalized abundance observed in the TRM10 strain (lane 3) set to 1. Triplicate data were plotted with each data point shown; error bars denote standard deviation. Two sample t-tests assuming equal variances were performed, and one-tailed P-values were shown with red asterisks under black bars for comparison of tRNATrp levels or a black asterisk or n.s. (no significant difference) over white bars for comparison of tRNAGly(GCC) levels. Specific P-values from other t-test comparisons are shown in Supplemental Table S1.
