c-Fos proto-oncoprotein is degraded by the proteasome independently of its own ubiquitinylation in vivo

Mol Cell Biol. 2003 Oct;23(20):7425-36. doi: 10.1128/MCB.23.20.7425-7436.2003.

Abstract

Prior ubiquitinylation of the unstable c-Fos proto-oncoprotein is thought to be required for recognition and degradation by the proteasome. Contradicting this view, we report that, although c-Fos can form conjugates with ubiquitin in vivo, nonubiquitinylatable c-Fos mutants show regulated degradation identical to that of the wild-type protein in living cells under two classical conditions of study: transient c-fos gene expression during the G(0)/G(1) phase transition upon stimulation by mitogens and constitutive expression during asynchronous growth. Moreover, c-Fos destruction during the G(0)/G(1) phase transition is unusual because it depends on two distinct but cumulative mechanisms. We report here that one mechanism involves a C-terminal destabilizer which does not need an active ubiquitin cycle, whereas the other involves an N-terminal destabilizer dependent on ubiquitinylation of an upstream c-Fos breakdown effector. In addition to providing new insights into the mechanisms of c-Fos protein destruction, an important consequence of our work is that ubiquitinylation-dependent proteasomal degradation claimed for a number of proteins should be reassessed on a new experimental basis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Division
  • Cysteine Endopeptidases / metabolism*
  • G1 Phase
  • HeLa Cells
  • Humans
  • Immunoblotting
  • Models, Biological
  • Multienzyme Complexes / metabolism*
  • Mutagenesis, Site-Directed
  • Phenotype
  • Plasmids / metabolism
  • Proteasome Endopeptidase Complex
  • Protein Binding
  • Protein Structure, Tertiary
  • Proto-Oncogene Proteins c-fos / metabolism*
  • Resting Phase, Cell Cycle
  • Temperature
  • Time Factors
  • Transcription, Genetic
  • Transfection
  • Ubiquitin / metabolism*

Substances

  • Multienzyme Complexes
  • Proto-Oncogene Proteins c-fos
  • Ubiquitin
  • Cysteine Endopeptidases
  • Proteasome Endopeptidase Complex