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Fig. 6. Drug sensitivities of scn1 and scn2 tRNA mutants, and high dosage suppression of scn mutants by Scn3 and Thr-tRNA. (A) Mutants scn1 and scn2 were hypersensitive to cycloheximide, a protein synthesis inhibitor. Wild-type, mis3-224 (Kondoh et al., 2000), scn1-17, scn2-7 and scn3+-disrupted cells (Samejima and Yanagida, 1994) were spotted on the YPD plate containing respective concentrations of cycloheximide. (B) Mutants scn1 and scn2 are hypersensitive to TBZ, a microtubule destabilizing drug at 26°C, the semi-permissive temperature. Wild-type, ß-tubulin mutant nda3-KM311 (Toda et al., 1983), scn1-17 and scn2-7 were spotted on the YPD plate containing respective concentrations of TBZ. (C) The scn3+ gene is a multicopy suppressor for scn1 and scn2 mutants. scn1-17 and scn2-7 mutants were transformed with plasmid containing the scn3+ gene under the inducible nmt1-promoter (pREP1-scn3+). Transformants were plated on the EMM2 containing thiamine (promoter, repressible condition). (D) Plasmid containing the Thr-tRNA(UGU) gene (pTRNAthr.06 and pTRNAthr.10) could suppress the cold-sensitivity of scn1 and scn2 mutant cells.





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