Assessing the importance of GCN2 regulation. Assays were conducted with wild type (wt), myo1Δ, gcn2Δ and myo1Δgcn2Δ strains. A) Western blot analysis of steady state levels of eIF2αp and its phosphorylated form eIF2α-P in total protein extracts derived from each strain. Pgk1p was used as a loading control, B) The morphological phenotype of each strain was observed by light microscopy at 100× magnification, C) Serial dilutions of cell suspensions from cultures of wild type (wt), myo1Δ, gcn2Δ, myo1Δgcn2Δ, and myo1Δgcn2ΔpRS316-MYO1+ strains were inoculated on CSM agar medium and allowed to grow for three days at 30°C for viability assays. The strain myo1Δgcn2ΔpRS316-MYO1+ was included to analyze complementation of the myo1Δgcn2Δ synthetic growth defect with a plasmid copy of the wild type MYO1 gene. Indicated at the top is the number of cells per 5 μL dilution.
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