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

A photoconvertible reporter of the ubiquitin-proteasome system in vivo.

Author information

1
Molecular Cancer Biology Program and Institute for Biomedicine, Biomedicum Helsinki, University of Helsinki, Helsinki, Finland.

Erratum in

  • Nat Methods. 2010 Jul;7(7):567.

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.

PMID:
20453865
DOI:
10.1038/nmeth.1460
[Indexed for MEDLINE]

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