Figure 3
From: Inactivation of NMD increases viability of sup45 nonsense mutants in Saccharomyces cerevisiae
Deletion of UPF1 gene leads to increased viability of sup45 nonsense mutants. (A). Strains 1A-D1628 (sup45Δ) and 1-1A-D1628 (sup45Δ upf1Δ) all containing SUP45 deletion and pRS316/SUP45 plasmid were transformed with pRS315/SUP45-LEU2 plasmids carrying different sup45 mutant alleles. The growth of the transformants was tested by plating 100, and 10-1 serial dilutions of overnight cultures (left to right) onto 5-FOA plates. The extent of cell growth on 5-FOA plates indicates the ability of the sup45 mutant alleles to support cell growth in the presence and absence of UPF1 gene. The same serially diluted cultures were also spotted on synthetic complete plates lacking leucine and uracil (-L -U) to estimate the total number of cells analyzed. (B). Level of eRF1 protein in the clones selected on 5-FOA medium was analyzed by western blot. Tubulin was used as a loading control. Following sup45 mutations were tested: 102, 107 (nonsense) and 103 (missense).