RRC ID 46601
Author Hamer G, Matilainen O, Holmberg CI.
Title A photoconvertible reporter of the ubiquitin-proteasome system in vivo.
Journal Nat Methods
Abstract 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.
Volume 7(6)
Pages 473-8
Published 2010-6-1
DOI 10.1038/nmeth.1460
PII nmeth.1460
PMID 20453865
MeSH Aging / metabolism Animals Caenorhabditis elegans / metabolism* Caenorhabditis elegans Proteins / physiology Dopamine / physiology Luminescent Proteins / metabolism* Microscopy, Fluorescence Proteasome Endopeptidase Complex / physiology* RNA Interference Receptors, Cytoplasmic and Nuclear / physiology Ubiquitin / metabolism* gamma-Aminobutyric Acid / physiology
IF 30.822
Times Cited 69
WOS Category BIOCHEMICAL RESEARCH METHODS
Resource
C.elegans tm1349