A trm140-Δ trm1Δ mutant is sensitive to cycloheximide. (A) Enhanced cycloheximide sensitivity is observed in a trm140-Δ trm1-Δ strain. Wild-type (BY4741), abp140-Δ (ySD179), trm1-Δ (yLN005), and abp140-Δ trm1-Δ (ySD576) strains were grown at 30°C overnight, and 0.3 OD cells were serially diluted by 10-fold, spotted on rich (YPD) medium containing either 0, 0.1, or 0.3 μg/mL cycloheximide, and incubated at 30°C for 2 d (without cycloheximide) or 3 d (with cycloheximide). (B) Cycloheximide growth inhibition of a trm140-Δ trm1Δ mutant is rescued by expression of TRM140 or TRM1. trm140-Δ trm1-Δ cells (ySD576) were transformed with a LEU2 CEN plasmid containing TRM140 (pSD158) or TRM1 (pMG67A) or with the control empty CEN LEU2 vector (AVA581), and transformants were grown overnight at 30°C in SD-Leu medium and spotted on YPD, YPG (glycerol) or YPD containing 0.25 μg/mL cycloheximide. Control wild-type (BY4741), trm140-Δ (ySD179), and trm1-Δ (yLN005) strains were transformed with the same empty vector and tested in parallel. Plates were incubated at 30°C for 2–3 d.
