Early-onset torsion dystonia: a novel high-throughput yeast genetic screen for factors modifying protein levels of torsinAΔE

• Published in: Disease models & mechanisms Vol. 10; no. 9; pp. 1129 - 1140
• Main Authors: Zacchi, Lucía F, Dittmar, John C, Mihalevic, Michael J, Shewan, Annette M, Schulz, Benjamin L, Brodsky, Jeffrey L, Bernstein, Kara A
• Format: Journal Article
• Language: English
• Published: England The Company of Biologists Ltd 01.09.2017; The Company of Biologists
• Subjects: Biosynthesis; Chemical bonds; Chromosomes; Disease; Dystonia; Dystonia Musculorum Deformans - genetics; Gene Ontology; Genes; Genes, Fungal; Genetic screen; Genetic Testing; Genomes; HeLa Cells; High-Throughput Screening Assays - methods; Humans; Library collections; Models, Biological; Molecular Chaperones - metabolism; Mutant Proteins - metabolism; Mutation; Protein disulfide isomerase; Protein Disulfide-Isomerases - metabolism; Protein levels; Protein Stability; Proteins; Quality control; Resource; Saccharomyces cerevisiae - genetics; TorsinA; Yeast; Protein disulfide isomerase; TorsinA; Protein levels; Yeast; Dystonia; Genetic screen
• ISSN: 1754-8403; 1754-8411
• DOI: 10.1242/dmm.029926

Dystonia is the third most common movement disorder, but its diagnosis and treatment remain challenging. One of the most severe types of dystonia is early-onset torsion dystonia (EOTD). The best studied and validated EOTD-associated mutation, torsinAΔE, is a deletion of a C-terminal glutamate residue in the AAA+ ATPase torsinA. TorsinA appears to be an endoplasmic reticulum (ER)/nuclear envelope chaperone with multiple roles in the secretory pathway and in determining subcellular architecture. Many functions are disabled in the torsinAΔE variant, and torsinAΔE is also less stable than wild-type torsinA and is a substrate for ER-associated degradation. Nevertheless, the molecular factors involved in the biogenesis and degradation of torsinA and torsinAΔE have not been fully explored. To identify conserved cellular factors that can alter torsinAΔE protein levels, we designed a new high-throughput, automated, genome-wide screen utilizing our validated torsinA expression system. By analyzing the yeast non-essential gene deletion collection, we identified 365 deletion strains with altered torsinAΔE steady-state levels. One notable hit was , which encodes a member of the protein disulfide isomerase family (PDIs). PDIs reside in the ER and catalyze the formation of disulfide bonds, mediate protein quality control and aid in nascent protein folding. We validated the role of select human PDIs in torsinA biogenesis in mammalian cells and found that overexpression of PDIs reduced the levels of torsinA and torsinAΔE. Together, our data report the first genome-wide screen to identify cellular factors that alter expression levels of the EOTD-associated protein torsinAΔE. More generally, the identified hits help in dissecting the cellular machinery involved in folding and degrading a torsinA variant, and constitute potential therapeutic factors for EOTD. This screen can also be readily adapted to identify factors impacting the levels of any protein of interest, considerably expanding the applicability of yeast in both basic and applied research.