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. 2018 Jan 12;19(3):e44867. doi: 10.15252/embr.201744867

Figure 3. Production and maturation of tRNA in hmt mutants.

Figure 3

  1. Aminoacylation of tRNA in nrs1/hmt1 and prs1/hmt2 mutants. Cells of the wild‐type (JY450), tor2‐ts6 (JV303), nrs1/hmt1 (JS159), and prs1/hmt2 (JS160) strains were grown in liquid YE medium at 25°C and then shifted to 36°C for 4 h. RNA extracted in acidic conditions was analyzed by northern blot analysis with probes for tRNA‐Asn and tRNA‐Pro. The leftmost and rightmost lanes are deacylated wild‐type RNA (DA). 5S rRNA is shown as a loading control.
  2. Amount of total tRNA in hmt mutants. Cells of the wild‐type (JY450), tor2‐ts6 (JV303), rpc34/hmt4 (JS162), and nrs1/hmt1 (JS159) strains were grown in liquid YE medium at 25°C and then shifted to 30°C for 4 h. Total RNA was separated, followed by ethidium bromide staining.
  3. Quantification of the amount of total tRNA in the wild‐type, tor2‐ts6, rpc34/hmt4, and nrs1/hmt1 cells. Data show the mean ± SD from three biological replicates, including the one in (B).
  4. Mating efficiency of the wild‐type (JY3) and maf1∆ (JS169) strains. Cells of each strain were incubated on SSA medium at 30°C for 3 days. Mean ± SD values of three independent measurements are shown (total n > 300).
  5. Mating efficiency of the wild‐type (JY3), tor2‐ts6 (JT360), tor2‐ts6 maf1∆ (JS170), and maf1∆ (JS169) strains on nutrient‐rich medium. Cells of each strain were incubated on YE medium at 30°C for 3 days. Mean ± SD values of three independent measurements are shown (total n > 300).