Prion-Dependent Lethality of Missense Mutations Is Caused by Low GTPase Activity of the Mutant eRF3 Protein.

The essential gene encodes yeast translation termination factor Sup35/eRF3. The N-terminal domain of Sup35 is also responsible for Sup35 prionization that leads to generation of the [] prion. Previously we isolated different types of mutations (missense and nonsense) and demonstrated that nonsense mutations () are incompatible with the [] prion, leading to lethality of [] haploid cells. Here, we show that missense mutations () within conservative regions of the Sup35 C-domain result in lethality of [] cells because of weak activity of Sup35/eRF3 as a translation termination factor. Mutant Sup35 maintain their ability to be incorporated into pre-existing [] aggregates and to form amyloid aggregates in vitro, while mutations do not influence the [] prion induction and stability. All these mutations (D363N, R372K, T378I) are located in the conservative GTPase region of Sup35, decreasing the GTPase activity of mutated proteins. We propose that such low activity of mutant Sup35 combined with aggregation of Sup35 constituting the [] prion is not sufficient to maintain the viability of yeast cells.