Cytomegalovirus promoter up-regulation is the major cause of increased protein levels of unstable reporter proteins after treatment of living cells with proteasome inhibitors

J Biol Chem. 2009 Oct 9;284(41):28253-28262. doi: 10.1074/jbc.M109.004101. Epub 2009 Aug 13.

Abstract

Fluorescent unstable proteins obtained by the fusion of a fluorescent protein coding sequence with specific amino acid sequences that promote its fast degradation have become popular to gauge the activity of the ubiquitin/proteasome system in living cells. The steady-state levels of expression of these unstable proteins is low in agreement with their short half-lives, and they accumulate in the cell upon treatment with proteasome inhibitors. We show here that this accumulation is mainly due to transcriptional up-regulation of the cytomegalovirus promoter by proteasome inhibitors and mediated, at least in part, by AP1 transactivation. These simple facts put under quarantine conclusions reached about the activity of the ubiquitin/proteasome pathway in animal cells in culture or in transgenic mice, where popular cytomegalovirus-driven constructs are used, as transcriptional regulation of the expression of the reporter protein construct and not degradation of the unstable protein by the ubiquitin/proteasome system may contribute significantly to the interpretation of the results observed.

Publication types

  • Research Support, Non-U.S. Gov't
  • Retracted Publication

MeSH terms

  • Animals
  • Cell Line
  • Cytomegalovirus* / drug effects
  • Cytomegalovirus* / genetics
  • Cytomegalovirus* / metabolism
  • Genes, Reporter
  • Humans
  • Mice
  • Mice, Transgenic
  • Promoter Regions, Genetic*
  • Protease Inhibitors / pharmacology*
  • Proteasome Endopeptidase Complex / metabolism
  • Proteasome Inhibitors*
  • Protein Biosynthesis
  • Rats
  • Recombinant Fusion Proteins* / genetics
  • Recombinant Fusion Proteins* / metabolism
  • Transcription, Genetic
  • Up-Regulation

Substances

  • Protease Inhibitors
  • Proteasome Inhibitors
  • Recombinant Fusion Proteins
  • Proteasome Endopeptidase Complex