p23 co-chaperone protects the aryl hydrocarbon receptor from degradation in mouse and human cell lines

Biochem Pharmacol. 2012 Sep 15;84(6):838-50. doi: 10.1016/j.bcp.2012.06.018. Epub 2012 Jul 1.

Abstract

The aryl hydrocarbon receptor (AhR) is a ligand-sensitive transcription factor which is responsible for most 2,3,7,8-tetrachlorodibenzo-p-dioxin toxicities. Without ligand, the AhR complex is cytoplasmic and contains p23. Our objective was to investigate whether the wild type p23 levels are important for the AhR function. We generated eight p23-specific knockdown stable cell lines via either electroporation or lentiviral infection. Five of these stable cell lines were generated from a mouse hepatoma cell line (Hepa1c1c7) and three were from human hepatoma and cervical cell lines (Hep3B and HeLa). All of them expressed lower AhR protein levels, leading to reduced ligand-induced, DRE-driven downstream activity. The AhR protein levels in p23-specific knockdown stable cells were reversed back to wild type levels after exogenous p23 was introduced. Reduction of the AhR protein levels in these stable cells was caused by a decrease in the AhR message levels and an increase of the AhR protein degradation in the absence of ligand. This ligand-independent degradation of AhR was not reversed by MG132, suggesting that the 26S proteasome was not responsible for the degradation. In addition, MG132 could not protect AhR from the ligand-induced degradation in both mouse and human p23-knockdown stable cells.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Cell Line, Tumor
  • Cytochrome P-450 CYP1A1 / genetics
  • Electroporation
  • Gene Knockdown Techniques
  • Humans
  • Hypoxia-Inducible Factor 1, alpha Subunit / metabolism
  • Intramolecular Oxidoreductases / genetics
  • Intramolecular Oxidoreductases / metabolism*
  • Lentivirus / genetics
  • Leupeptins / pharmacology
  • Methylcholanthrene / pharmacology
  • Mice
  • Polychlorinated Dibenzodioxins / toxicity
  • Promoter Regions, Genetic
  • Prostaglandin-E Synthases
  • Receptors, Aryl Hydrocarbon / agonists
  • Receptors, Aryl Hydrocarbon / antagonists & inhibitors
  • Receptors, Aryl Hydrocarbon / metabolism*
  • Response Elements

Substances

  • Hypoxia-Inducible Factor 1, alpha Subunit
  • Leupeptins
  • Polychlorinated Dibenzodioxins
  • Receptors, Aryl Hydrocarbon
  • Methylcholanthrene
  • Cytochrome P-450 CYP1A1
  • Intramolecular Oxidoreductases
  • Prostaglandin-E Synthases
  • Ptges3 protein, mouse
  • benzyloxycarbonylleucyl-leucyl-leucine aldehyde