Sterol-regulated degradation of Insig-1 mediated by the membrane-bound ubiquitin ligase gp78

J Biol Chem. 2006 Dec 22;281(51):39308-15. doi: 10.1074/jbc.M608999200. Epub 2006 Oct 16.

Abstract

Insig-1 and Insig-2, closely related endoplasmic reticulum membrane proteins, mediate transcriptional and post-transcriptional mechanisms that assure cholesterol homeostasis through their sterol-induced binding to Scap (SREBP cleavage-activating protein) and 3-hydroxy-3-methylglutaryl coenzyme A reductase. Recent studies show that Insig-1 (but not Insig-2) is ubiquitinated and rapidly degraded when cells are depleted of sterols. Conversely, ubiquitination of Insig-1 is blocked, and the protein is stabilized when intracellular sterols accumulate. Here, we report that the ubiquitin ligase gp78, which binds with much higher affinity to Insig-1 than Insig-2, is required for ubiquitination and degradation of Insig-1 in sterol-depleted cells. Sterols prevent Insig-1 ubiquitination and degradation by displacing gp78 from Insig-1, an event that results from sterol-induced binding of Scap to Insig-1. In addition to providing a mechanism for sterol-regulated degradation of Insig-1, these results help to explain why Scap is subject to endoplasmic reticulum retention upon Insig-1 binding, whereas 3-hydroxy-3-methylglutaryl coenzyme A reductase is ubiquitinated and degraded.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line
  • Cell Membrane / metabolism*
  • Endoplasmic Reticulum / metabolism
  • Humans
  • Hydroxymethylglutaryl CoA Reductases / metabolism
  • Intracellular Signaling Peptides and Proteins / metabolism
  • Membrane Proteins / metabolism*
  • Models, Biological
  • Plasmids / metabolism
  • Protein Binding
  • RNA Interference
  • Receptors, Autocrine Motility Factor
  • Receptors, Cytokine / chemistry
  • Receptors, Cytokine / physiology*
  • Sterols / metabolism*
  • Ubiquitin / metabolism
  • Ubiquitin-Protein Ligases / chemistry
  • Ubiquitin-Protein Ligases / physiology*

Substances

  • INSIG1 protein, human
  • INSIG2 protein, human
  • Intracellular Signaling Peptides and Proteins
  • Membrane Proteins
  • Receptors, Cytokine
  • SREBP cleavage-activating protein
  • Sterols
  • Ubiquitin
  • Hydroxymethylglutaryl CoA Reductases
  • AMFR protein, human
  • Receptors, Autocrine Motility Factor
  • Ubiquitin-Protein Ligases