Figure 2.

Hyperactive eRF1 mutants can complement the deletion of the SUP45 gene. The FS1ΔS strain was transformed with a plasmid carrying sup45 mutants and plated on selective media for this plasmid. Rescue plasmid carrying wild-type SUP45 gene was shuffled by plating the colonies on media containing 5FOA. The growing cells were transferred to a selective liquid culture until they reached 1 OD₆₀₀ and were serially diluted. Plates were incubated either at 30 °C or 37 °C to test for thermosensitivity. Ts denotes the MT557/3b strain carrying a thermosensitive sup45 allele; E104K, E360G; Q76R, E360G; N58K, E360G; are double mutants created by directed mutagenesis, combining mutations identified independently during the screen within the same molecule.