Fig 3. Spontaneous suppressors of S. pombe trm6Δ mutants with mutations in dhp1 and tol1 restore growth and increase tRNAiMet(CAU) levels of S. pombe trm6Δ mutants.
(A) Spontaneous suppressors of S. pombe trm6Δ mutants with mutations in dhp1 and tol1 restore growth at high temperatures. Strains as indicated were grown overnight in YES media at 30°C and analyzed for growth as in Fig 1A on indicated plates and temperatures. (B) Spontaneous suppressors of S. pombe trm6Δ mutants with mutations in dhp1 and tol1 restore tRNAiMet(CAU) levels after growth at 38.5°C. Strains were grown in YES media at 30°C and shifted to 38.5°C for 8 hours as described in Materials and Methods, and RNA was isolated and analyzed by northern blotting as in Fig 2A. (C) Quantification of tRNA levels of S. pombe trm6Δ dhp1 and tol1 mutants. tRNA levels were quantified as in Fig 2B, with dark and light shades as indicated for 30°C and 38.5°C.