泛素
秀丽隐杆线虫
蛋白酶体
细胞生物学
绿色荧光蛋白
报告基因
蛋白质降解
突变体
融合蛋白
生物
转基因
化学
生物化学
基因
基因表达
重组DNA
作者
Geert Hamer,Olli Matilainen,Carina I. Holmberg
出处
期刊:Nature Methods
[Springer Nature]
日期:2010-05-09
卷期号:7 (6): 473-478
被引量:104
摘要
A photoconvertible reporter of the ubiquitin-proteasome system permits detection of its activity independent of protein synthesis and is applied to study cell type– and age-specific protein degradation in living Caenorhabditis elegans. The ubiquitin-proteasome system (UPS) orchestrates many cellular and tissue-specific processes by degrading damaged and key regulatory proteins. To enable investigation of UPS activity in different cell types in a living animal, we developed a photoconvertible fluorescent UPS reporter system for live imaging and quantification of protein degradation in Caenorhabditis elegans. Our reporter consists of the photoconvertible fluorescent protein Dendra2 targeted for proteasomal degradation by fusion to the UbG76V mutant form of ubiquitin. In contrast to previous reporters, this system permits quantification of UPS activity independently of protein synthesis. Our reporter revealed that UPS-mediated protein degradation varies in a cell type–specific and age-dependent manner in C. elegans.
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