A photoconvertible reporter of the ubiquitin-proteasome system in vivo.
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.
Hamer
Geert
G
Molecular Cancer Biology Program and Institute for Biomedicine, Biomedicum Helsinki, University of Helsinki, Helsinki, Finland.
Matilainen
Olli
O
Holmberg
Carina I
CI
D000375:Aging / Q000378:metabolism; D000818:Animals; D017173:Caenorhabditis elegans / Q000378:metabolism*; D029742:Caenorhabditis elegans Proteins / Q000502:physiology; D004298:Dopamine / Q000502:physiology; D008164:Luminescent Proteins / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D046988:Proteasome Endopeptidase Complex / Q000502:physiology*; D034622:RNA Interference; D018160:Receptors, Cytoplasmic and Nuclear / Q000502:physiology; D025801:Ubiquitin / Q000378:metabolism*; D005680:gamma-Aminobutyric Acid / Q000502:physiology
D029742:Caenorhabditis elegans Proteins; D008164:Luminescent Proteins; C516341:RPN-10 protein, C elegans; D018160:Receptors, Cytoplasmic and Nuclear; D025801:Ubiquitin; D005680:gamma-Aminobutyric Acid; D046988:Proteasome Endopeptidase Complex; D004298:Dopamine
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