Dopamine D(3) receptors are down-regulated following heterologous endocytosis by a specific interaction with G protein-coupled receptor-associated sorting protein-1

Dawn Thompson, Jennifer L Whistler

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

The D(3) dopamine receptor is endocytosed through a heterologous mechanism mediated by phorbol esters. Here, we show that following this endocytosis the D(3) 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 D(2) that confers GASP-1 binding. shRNA knockdown of GASP-1 delayed post-endocytic degradation of both the D(2) and D(3) dopamine receptors. In addition, mutation of the D(2) and D(3) receptor C termini to resemble the D(4), 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 D(4) to resemble that of the D(2) and D(3) facilitated GASP-1 binding and promoted post-endocytic degradation of the mutant D(4) 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
Pages (from-to)1598-608
Number of pages11
JournalThe Journal of Biological Chemistry
Volume286
Issue number2
DOIs
Publication statusPublished - 14 Jan 2011

Keywords

  • Amino Acid Sequence
  • Down-Regulation
  • Endocytosis
  • HEK293 Cells
  • Humans
  • Molecular Sequence Data
  • Mutagenesis
  • Phorbol Esters
  • Protein Binding
  • Protein Structure, Tertiary
  • Protein Transport
  • Proteins
  • RNA, Small Interfering
  • Receptors, Dopamine D2
  • Receptors, Dopamine D3
  • Receptors, Dopamine D4

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