The Mof2/Sui1 Protein Is a General Monitor of Translational Accuracy

ABSTRACT

Although it is essential for protein synthesis to be highly accurate, a number of cases of directed ribosomal frameshifting have been reported in RNA viruses, as well as in procaryotic and eucaryotic genes. Changes in the efficiency of ribosomal frameshifting can have major effects on the ability of cells to propagate viruses which use this mechanism. Furthermore, studies of this process can illuminate the mechanisms involved in the maintenance of the normal translation reading frame. The yeast Saccharomyces cerevisiae killer virus system uses programmed −1 ribosomal frameshifting to synthesize its gene products. Strains harboring the mof2-1 allele demonstrated a fivefold increase in frameshifting and prevented killer virus propagation. In this report, we present the results of the cloning and characterization of the wild-type MOF2 gene.mof2-1 is a novel allele of SUI1, a gene previously shown to play a role in translation initiation start site selection. Strains harboring the mof2-1 allele demonstrated a mutant start site selection phenotype and increased efficiency of programmed −1 ribosomal frameshifting and conferred paromomycin sensitivity. The increased frameshifting observed in vivo was reproduced in extracts prepared from mof2-1 cells. Addition of purified wild-type Mof2p/Sui1p reduced frameshifting efficiencies to wild-type levels. Expression of the human SUI1 homolog in yeast corrects all of the mof2-1 phenotypes, demonstrating that the function of this protein is conserved throughout evolution. Taken together, these results suggest that Mof2p/Sui1p functions as a general modulator of accuracy at both the initiation and elongation phases of translation.

ACKNOWLEDGMENTS

This work was supported by a grant (GM48631) from the National Institutes of Health and an American Heart Association Established Investigator Award to S.W.P. and by grant 8-97 to J.D.D. from the UMDNJ Foundation.

We thank Carlos Gonzalez, Maria Ruiz-Echevarria, Shuang Zhang, Kevin Czaplinski, and Philip Farabaugh for discussions of the work and critical reading of the manuscript. We thank Thomas Donahue for sending us strains and plasmids and Alan Hinnebusch for strains harboring mutations in the GCD10 gene.