A yeast selection system for the detection of proteasomal activation

The ubiquitin proteasome system (UPS) is a complex cellular machinery that catalyzes degradation of misfolded or damaged proteins and regulates turnover of native proteins in eukaryotic cells, thus playing a crucial role in maintaining protein homeostasis. The UPS has emerged as a drug target for a diverse range of diseases characterized by accumulation of misfolded or aggregated proteins. While enhancement of UPS activity is widely recognized as a promising strategy to prevent accumulation of aberrant, off-pathway protein conformations and ameliorate the phenotypes of a wide range of protein misfolding diseases, the molecular mechanisms underlying activation of proteasomal degradation are poorly characterized. We report the development of a yeast selection platform for genome-wide selection of UPS activators. We engineered the Saccharomyces cerevisiae selection marker orotidine-5'-phosphate decarboxylase (URA3) to function as a substrate of proteasomal degradation through fusion to UPS-sensitive tags. The resulting UPS-sensitive URA3 variant links UPS activity to cell growth. The yeast selection platform reported in this study will open the way to high-throughput, genome-wide studies aimed at identifying modulators of UPS function that might provide novel target for therapeutic applications.