A Genetic Tool to Track Protein Aggregates and Control Prion Inheritance

• Published in: Cell 2017-11, Vol.171 (4), p.966-979.e18
• Main Authors: Newby, Gregory A., Kiriakov, Szilvia, Hallacli, Erinc, Kayatekin, Can, Tsvetkov, Peter, Mancuso, Christopher P., Bonner, J. Maeve, Hesse, William R., Chakrabortee, Sohini, Manogaran, Anita L., Liebman, Susan W., Lindquist, Susan, Khalil, Ahmad S.
• Format: Article
• Language: English
• Subjects: epigenetics; Genetic Engineering; Genetic Techniques - economics; high-throughput screening; memory; mRNA Cleavage and Polyadenylation Factors - metabolism; prions; Prions - genetics; protein aggregation; RNA-binding proteins; RNA-Binding Proteins - metabolism; Saccharomyces cerevisiae - genetics; Saccharomyces cerevisiae Proteins - metabolism; synthetic biology; Synthetic Biology - methods; synthetic transcription factor; yTRAP
• ISSN: 0092-8674; 1097-4172
• DOI: 10.1016/j.cell.2017.09.041

Protein aggregation is a hallmark of many diseases but also underlies a wide range of positive cellular functions. This phenomenon has been difficult to study because of a lack of quantitative and high-throughput cellular tools. Here, we develop a synthetic genetic tool to sense and control protein aggregation. We apply the technology to yeast prions, developing sensors to track their aggregation states and employing prion fusions to encode synthetic memories in yeast cells. Utilizing high-throughput screens, we identify prion-curing mutants and engineer “anti-prion drives” that reverse the non-Mendelian inheritance pattern of prions and eliminate them from yeast populations. We extend our technology to yeast RNA-binding proteins (RBPs) by tracking their propensity to aggregate, searching for co-occurring aggregates, and uncovering a group of coalescing RBPs through screens enabled by our platform. Our work establishes a quantitative, high-throughput, and generalizable technology to study and control diverse protein aggregation processes in cells. [Display omitted] •yTRAP is a platform for quantitative in vivo analysis of protein aggregation•High-throughput screens uncover mutant prion-curing alleles•Synthetic memory and “anti-prion drives” are engineered with prion-switching tools•Screens reveal aggregation-prone RNA-binding proteins and their interactors Newby et al. developed a new tool for tracking and quantifying protein aggregation and prion inheritance in live cells.