The Deubiquitylase USP2 Regulates the LDLR Pathway by Counteracting the E3-Ubiquitin Ligase IDOL

Circ Res. 2016 Feb 5;118(3):410-9. doi: 10.1161/CIRCRESAHA.115.307298. Epub 2015 Dec 14.

Abstract

Rationale: The low-density lipoprotein (LDL) receptor (LDLR) is a central determinant of circulating LDL-cholesterol and as such subject to tight regulation. Recent studies and genetic evidence implicate the inducible degrader of the LDLR (IDOL) as a regulator of LDLR abundance and of circulating levels of LDL-cholesterol in humans. Acting as an E3-ubiquitin ligase, IDOL promotes ubiquitylation and subsequent lysosomal degradation of the LDLR. Consequently, inhibition of IDOL-mediated degradation of the LDLR represents a potential strategy to increase hepatic LDL-cholesterol clearance.

Objective: To establish whether deubiquitylases counteract IDOL-mediated ubiquitylation and degradation of the LDLR.

Methods and results: Using a genetic screening approach, we identify the ubiquitin-specific protease 2 (USP2) as a post-transcriptional regulator of IDOL-mediated LDLR degradation. We demonstrate that both USP2 isoforms, USP2-69 and USP2-45, interact with IDOL and promote its deubiquitylation. IDOL deubiquitylation requires USP2 enzymatic activity and leads to a marked stabilization of IDOL protein. Paradoxically, this also markedly attenuates IDOL-mediated degradation of the LDLR and the ability of IDOL to limit LDL uptake into cells. Conversely, loss of USP2 reduces LDLR protein in an IDOL-dependent manner and limits LDL uptake. We identify a tri-partite complex encompassing IDOL, USP2, and LDLR and demonstrate that in this context USP2 promotes deubiquitylation of the LDLR and prevents its degradation.

Conclusions: Our findings identify USP2 as a novel regulator of lipoprotein clearance owing to its ability to control ubiquitylation-dependent degradation of the LDLR by IDOL.

Keywords: E3 ubiquitin ligase; LDL cholesterol; LDL receptors; MYLIP/IDOL; USP2; ubiquitin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cholesterol, LDL / metabolism*
  • Endopeptidases / genetics
  • Endopeptidases / metabolism*
  • Enzyme Stability
  • HEK293 Cells
  • HeLa Cells
  • Hep G2 Cells
  • Humans
  • Mice, Knockout
  • Multienzyme Complexes
  • Protein Binding
  • Proteolysis
  • RNA Interference
  • Receptors, LDL / genetics
  • Receptors, LDL / metabolism*
  • Transfection
  • Ubiquitin Thiolesterase
  • Ubiquitin-Protein Ligases / deficiency
  • Ubiquitin-Protein Ligases / genetics
  • Ubiquitin-Protein Ligases / metabolism*
  • Ubiquitination

Substances

  • Cholesterol, LDL
  • LDLR protein, human
  • Multienzyme Complexes
  • Receptors, LDL
  • MYLIP protein, human
  • Mylip protein, mouse
  • Ubiquitin-Protein Ligases
  • Endopeptidases
  • USP2 protein, human
  • Ubiquitin Thiolesterase