Both exonucleolytic 3′ end maturation pathway and chaperone function of Lhp1 influence the precursor pattern of tested tRNAs. (A–C) Wild-type and indicated mutant strains were grown at 30°C to exponential phase and then shifted for 2 h to 30°C or 37°C and/or YPGly. RNA was isolated and analyzed by Northern hybridization with probes specific to tRNAUAUIle, tRNAUUULys, and tRNATyr. Ethidium bromide–stained 5.8S rRNA was used as a loading control. The major and minor forms of pre-tRNA precursors that accumulate under stress conditions are indicated by solid and dashed arrows, respectively. (D) Calculation of ratios between various forms of pre-tRNA (specified in the legend to Fig. 1) from at least three biological replicates.
