Dopamine D3 receptors are down-regulated following heterologous endocytosis by a specific interaction with G protein-coupled receptor-associated sorting protein-1

Dawn Thompson, Jennifer Whistler

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

The D3 dopamine receptor is endocytosed through a heterologous mechanism mediated by phorbol esters. Here, we show that following this endocytosis the D3 dopamine receptors fail to recycle and are instead targeted for degradation through an interaction with the G protein-coupled receptor (GPCR)-associated sorting protein-1 (GASP-1). Furthermore, we identified a specific binding motif in the C terminus common to the D 3 and D2 that confers GASP-1 binding. shRNA knockdown of GASP-1 delayed post-endocytic degradation of both the D2 and D 3 dopamine receptors. In addition, mutation of the D2 and D3 receptor C termini to resemble the D4, which does not interact with GASP-1, not only inhibited GASP-1 binding but slowed degradation after endocytosis. Conversely, mutation of the C terminus of the D4 to resemble that of the D2 and D3 facilitated GASP-1 binding and promoted post-endocytic degradation of the mutant D4 receptor. Thus, we have identified a motif that is both necessary and sufficient to promote GASP-1 binding and receptor degradation. In addition, these data demonstrated that GASP-1 can mediate post-endocytic degradation of dopamine receptors that have been endocytosed not only as a consequence of dopamine activation but also as a consequence of activation by phorbol esters.

Original languageEnglish (US)
Pages (from-to)1598-1608
Number of pages11
JournalJournal of Biological Chemistry
Volume286
Issue number2
DOIs
StatePublished - Jan 14 2011
Externally publishedYes

Fingerprint

Dopamine D3 Receptors
Protein Transport
G-Protein-Coupled Receptors
Endocytosis
Sorting
Protein Binding
Dopamine Receptors
Phorbol Esters
Degradation
Proteins
Mutation
Small Interfering RNA
Dopamine
Chemical activation

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

@article{689fbb2d69eb4a3ea3b1405233f1ffaf,
title = "Dopamine D3 receptors are down-regulated following heterologous endocytosis by a specific interaction with G protein-coupled receptor-associated sorting protein-1",
abstract = "The D3 dopamine receptor is endocytosed through a heterologous mechanism mediated by phorbol esters. Here, we show that following this endocytosis the D3 dopamine receptors fail to recycle and are instead targeted for degradation through an interaction with the G protein-coupled receptor (GPCR)-associated sorting protein-1 (GASP-1). Furthermore, we identified a specific binding motif in the C terminus common to the D 3 and D2 that confers GASP-1 binding. shRNA knockdown of GASP-1 delayed post-endocytic degradation of both the D2 and D 3 dopamine receptors. In addition, mutation of the D2 and D3 receptor C termini to resemble the D4, which does not interact with GASP-1, not only inhibited GASP-1 binding but slowed degradation after endocytosis. Conversely, mutation of the C terminus of the D4 to resemble that of the D2 and D3 facilitated GASP-1 binding and promoted post-endocytic degradation of the mutant D4 receptor. Thus, we have identified a motif that is both necessary and sufficient to promote GASP-1 binding and receptor degradation. In addition, these data demonstrated that GASP-1 can mediate post-endocytic degradation of dopamine receptors that have been endocytosed not only as a consequence of dopamine activation but also as a consequence of activation by phorbol esters.",
author = "Dawn Thompson and Jennifer Whistler",
year = "2011",
month = "1",
day = "14",
doi = "10.1074/jbc.M110.158345",
language = "English (US)",
volume = "286",
pages = "1598--1608",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "2",

}

TY - JOUR

T1 - Dopamine D3 receptors are down-regulated following heterologous endocytosis by a specific interaction with G protein-coupled receptor-associated sorting protein-1

AU - Thompson, Dawn

AU - Whistler, Jennifer

PY - 2011/1/14

Y1 - 2011/1/14

N2 - The D3 dopamine receptor is endocytosed through a heterologous mechanism mediated by phorbol esters. Here, we show that following this endocytosis the D3 dopamine receptors fail to recycle and are instead targeted for degradation through an interaction with the G protein-coupled receptor (GPCR)-associated sorting protein-1 (GASP-1). Furthermore, we identified a specific binding motif in the C terminus common to the D 3 and D2 that confers GASP-1 binding. shRNA knockdown of GASP-1 delayed post-endocytic degradation of both the D2 and D 3 dopamine receptors. In addition, mutation of the D2 and D3 receptor C termini to resemble the D4, which does not interact with GASP-1, not only inhibited GASP-1 binding but slowed degradation after endocytosis. Conversely, mutation of the C terminus of the D4 to resemble that of the D2 and D3 facilitated GASP-1 binding and promoted post-endocytic degradation of the mutant D4 receptor. Thus, we have identified a motif that is both necessary and sufficient to promote GASP-1 binding and receptor degradation. In addition, these data demonstrated that GASP-1 can mediate post-endocytic degradation of dopamine receptors that have been endocytosed not only as a consequence of dopamine activation but also as a consequence of activation by phorbol esters.

AB - The D3 dopamine receptor is endocytosed through a heterologous mechanism mediated by phorbol esters. Here, we show that following this endocytosis the D3 dopamine receptors fail to recycle and are instead targeted for degradation through an interaction with the G protein-coupled receptor (GPCR)-associated sorting protein-1 (GASP-1). Furthermore, we identified a specific binding motif in the C terminus common to the D 3 and D2 that confers GASP-1 binding. shRNA knockdown of GASP-1 delayed post-endocytic degradation of both the D2 and D 3 dopamine receptors. In addition, mutation of the D2 and D3 receptor C termini to resemble the D4, which does not interact with GASP-1, not only inhibited GASP-1 binding but slowed degradation after endocytosis. Conversely, mutation of the C terminus of the D4 to resemble that of the D2 and D3 facilitated GASP-1 binding and promoted post-endocytic degradation of the mutant D4 receptor. Thus, we have identified a motif that is both necessary and sufficient to promote GASP-1 binding and receptor degradation. In addition, these data demonstrated that GASP-1 can mediate post-endocytic degradation of dopamine receptors that have been endocytosed not only as a consequence of dopamine activation but also as a consequence of activation by phorbol esters.

UR - http://www.scopus.com/inward/record.url?scp=78651399026&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=78651399026&partnerID=8YFLogxK

U2 - 10.1074/jbc.M110.158345

DO - 10.1074/jbc.M110.158345

M3 - Article

VL - 286

SP - 1598

EP - 1608

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 2

ER -