A photoconvertible reporter of the ubiquitin-proteasome system in vivo

Nat Methods. 2010 Jun;7(6):473-8. doi: 10.1038/nmeth.1460. Epub 2010 May 9.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 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

Substances

  • Caenorhabditis elegans Proteins
  • Luminescent Proteins
  • RPN-10 protein, C elegans
  • Receptors, Cytoplasmic and Nuclear
  • Ubiquitin
  • gamma-Aminobutyric Acid
  • Proteasome Endopeptidase Complex
  • Dopamine