ER stress induces alternative nonproteasomal degradation of ER proteins but not of cytosolic ones

Cell Stress Chaperones. Winter 2007;12(4):373-83. doi: 10.1379/csc-281.1.


Inhibition of protein folding in the endoplasmic reticulum (ER) causes ER stress, which triggers the unfolded protein response (UPR). To decrease the biosynthetic burden on the ER, the UPR inhibits in its initial stages protein synthesis. At later stages it upregulates components of ER-associated degradation (ERAD) and of the ubiquitin/proteasome system, which targets ER as well as cytosolic proteins for disposal. Here we report that, at later stages, the UPR also activates an alternative nonproteasomal pathway of degradation, which is resistant to proteasome inhibitors and is specific for ER substrates (assessed with uncleaved precursor of asialoglycoprotein receptor H2a and unassembled CD3delta) and not for cytosolic ones (p53). To mimic the initial inhibition of translation during UPR, we incubated cells with cycloheximide. After this treatment, degradation of ERAD substrates was no longer effected by proteasomal inhibition, similarly to the observed outcome of UPR. The degradation also became insensitive to abrogation of ubiquitination in a cell line carrying a thermosensitive E1 ubiquitin activating enzyme mutant. Of all protease inhibitors tested, only the metal chelator o-phenanthroline could block this nonproteasomal degradation. Preincubation of o-phenanthroline with Mn2+ or Co2+, but not with other cations, reversed the inhibition. Our results suggest that, upon inhibition of translation, an alternative nonproteasomal pathway is activated for degradation of proteins from the ER. This involves a Mn2+/Co2+-dependent metalloprotease or other metalloprotein. The alternative pathway selectively targets ERAD substrates to reduce the ER burden, but does not affect p53, the levels of which remain dependent on proteasomal control.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CHO Cells
  • Cobalt
  • Cricetinae
  • Cricetulus
  • Cycloheximide / pharmacology
  • Cytosol / drug effects
  • Cytosol / metabolism
  • Endoplasmic Reticulum / drug effects
  • Endoplasmic Reticulum / metabolism*
  • Endoplasmic Reticulum / pathology
  • Manganese
  • Metalloproteins / metabolism
  • Mice
  • Models, Biological
  • Mutation / genetics
  • NIH 3T3 Cells
  • Phenanthrolines / pharmacology
  • Proteasome Endopeptidase Complex / metabolism
  • Proteasome Inhibitors
  • Protein Biosynthesis / drug effects
  • Protein Folding*
  • Protein Processing, Post-Translational* / drug effects
  • Proteins / metabolism*
  • Temperature
  • Tumor Suppressor Protein p53 / metabolism
  • Ubiquitination / drug effects


  • Metalloproteins
  • Phenanthrolines
  • Proteasome Inhibitors
  • Proteins
  • Tumor Suppressor Protein p53
  • Cobalt
  • Manganese
  • Cycloheximide
  • Proteasome Endopeptidase Complex