Fig 5. Spontaneous suppressors of S. pombe trm6Δ imt06Δ mutants with mutations in dhp1 and tol1 restore growth and increase tRNAiMet(CAU) levels.
(A) Spontaneous suppressors of S. pombe trm6Δ imt06Δ mutants with mutations in dhp1 and tol1 suppress the growth defect. Strains 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 imt06Δ mutants with mutations in dhp1 and tol1 restore tRNAiMet(CAU) levels at low and high temperatures. Strains were grown in YES media at 30°C and shifted to 38.5°C for 6 hours as described in Materials and Methods, and RNA was isolated and analyzed by northern blotting as in Fig 2A. (C) Quantification of tRNAiMet(CAU) levels in S. pombe trm6Δ imt06Δ dhp1-7 and trm6Δ imt06Δ tol1-2 mutants. tRNA levels were quantified as in Fig 2B.