Figure 6
From: Inactivation of NMD increases viability of sup45 nonsense mutants in Saccharomyces cerevisiae
Deletion either UPF2 or UPF3 gene leads to increased viability of sup45 nonsense mutants. Strains 4a-D1659 (sup45Δ) and 4b-D1659 (sup45Δ upf2Δ) (A), 18a-D1660 (sup45Δ) and 3a-D1660 (sup45Δ upf3Δ) (B), all containing SUP45 deletion and pRS316/SUP45 plasmid were transformed with pRS315/sup45-n-LEU2 plasmids carrying different sup45 mutant alleles. The growth of the transformants was tested by plating 100, and 101 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 UPF2 or UPF3 genes. The same serially diluted cultures were also spotted on synthetic complete plates lacking leucine and uracil to estimate the total number of cells analyzed. (C, D) eRF1 and eRF3 protein levels in the same transformants as in panels A and B were analyzed by western blot. Tubulin was used as a loading control. Following sup45 mutations were tested: 102, 107 (nonsense) and 103 (missense).