A yeast selection system for the detection of proteasomal activation
A yeast selection system for the detection of proteasomal activation
复制标题
DOI：
10.1093/protein/gzz006
复制
发表时间：
2018-11-01
影响因子：
2.4
通讯作者：
Segatori，Laura
中科院分区：
文献类型：
--
作者：
Zhao，Wenting;Bachhav，Bhagyashree;Segatori，Laura
泛素蛋白酶体系统（ubiquitin proteasome system，UPS）是真核细胞中一种复杂的细胞机制，催化错误折叠或受损蛋白质的降解，调节天然蛋白质的周转，在维持蛋白质稳态中起着至关重要的作用。UPS已成为以错误折叠或聚集蛋白质积累为特征的多种疾病的药物靶标。虽然UPS活性的增强被广泛认为是一种有前途的策略，以防止异常的，关闭途径的蛋白质构象的积累和改善广泛的蛋白质错误折叠疾病的表型，蛋白酶体降解的激活的分子机制的特点是很差的。我们报告了一个酵母选择平台的开发，用于UPS激活剂的全基因组选择。我们将酵母选择标记乳清酸核苷-5 ′-磷酸脱羧酶（orotidine-5′-phosphate decarboxylase，URA 3）与UPS敏感标签融合，使其成为蛋白酶体降解的底物。由此产生的UPS敏感的URA 3变体将UPS活性与细胞生长联系起来。本研究中报道的酵母选择平台将为高通量、全基因组研究开辟道路，这些研究旨在鉴定可能为治疗应用提供新靶点的UPS功能调节剂。
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 theSaccharomyces cerevisiaeselection 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.