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Dopamine D4 receptor ubiquitination.

Kamila Skieterska1, Pieter Rondou2, Kathleen Van Craenenbroeck3

  • 1Laboratory of GPCR Expression and Signal Transduction (L-GEST), Ghent University, Proeftuinstraat 86, 9000 Gent, Belgium.

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|April 13, 2016
PubMed
Summary

The protein KLHL12 enhances ubiquitination of the dopamine D4 receptor (D4R) variants, particularly those with two or four repeats, suggesting functional implications for attention deficit hyperactivity disorder (ADHD) risk. Ubiquitination occurs on non-lysine residues and does not lead to D4R degradation.

Keywords:
G protein-coupled receptor (GPCR)KLHL12dopamine D4 receptornon-lysine ubiquitinationubiquitination

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Neuroscience

Background:

  • Ubiquitination is a key post-translational modification regulating protein fate and cellular functions.
  • The dopamine D4 receptor (D4R), a G protein-coupled receptor (GPCR), exhibits polymorphic variants in its third intracellular loop, linked to attention deficit hyperactivity disorder (ADHD).

Purpose of the Study:

  • To investigate the ubiquitination of dopamine D4 receptor (D4R) variants.
  • To identify proteins interacting with the polymorphic region of D4R and their role in ubiquitination.
  • To determine the functional consequences of D4R ubiquitination by KLHL12.

Main Methods:

  • Co-immunoprecipitation to identify interacting proteins.
  • Western blotting to assess ubiquitination levels of D4R variants.
  • Site-directed mutagenesis to investigate ubiquitination sites.

Main Results:

  • KLHL12 specifically interacts with the polymorphic region of D4R and enhances its ubiquitination.
  • KLHL12 strongly promotes ubiquitination of two- and four-repeat D4R variants, but not the seven-repeat variant.
  • D4R ubiquitination by KLHL12 occurs on non-lysine residues (cysteine, serine/threonine) and does not result in receptor degradation.

Conclusions:

  • Differential ubiquitination of D4R polymorphic variants by KLHL12 may have functional implications.
  • D4R can be ubiquitinated on non-lysine residues, expanding the understanding of ubiquitination mechanisms in GPCRs.
  • KLHL12-mediated ubiquitination influences D4R signaling or trafficking rather than degradation.