The G-protein coupled receptor associated sorting protein GASP-1 regulates the signalling and trafficking of the viral chemokine receptor US28

Traffic. 2010 May;11(5):660-74. doi: 10.1111/j.1600-0854.2010.1045.x.

Abstract

Human cytomegalovirus (HCMV) encodes the seven transmembrane(7TM)/G-protein coupled receptor (GPCR) US28, which signals and endocytoses in a constitutive, ligand-independent manner. Here we show that, following endocytosis, US28 is targeted to the lysosomes for degradation as a consequence of its interaction with the GPCR-associated sorting protein-1 (GASP-1). We find that GASP-1 binds to US28 in vitro and that disruption of the GASP-1/US28 interaction by either (i) overexpression of dominant negative cGASP-1 or by (ii) shRNA knock-down of endogenous GASP-1 is sufficient to inhibit the lysosomal targeting of US28 and slow its post-endocytic degradation. Furthermore, we found that GASP-1 affects US28-mediated signalling. The knock-down of endogenous GASP-1 impairs the US28-mediated Galphaq/PLC/inositol phosphate (IP) accumulation as well as the activation of the transcription factors Nuclear Factor-kappaB (NF-kappaB) and cyclic AMP responsive element binding protein (CREB). Overexpression of GASP-1 enhances both IP accumulation and transcription factor activity. Thus, GASP-1 is an important cellular determinant that not only regulates the post-endocytic trafficking of US28, but also regulates the signalling capacities of US28.

Publication types

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

MeSH terms

  • Chemokines / metabolism
  • Cyclic AMP Response Element-Binding Protein / metabolism
  • Cytomegalovirus / genetics
  • Cytomegalovirus / metabolism
  • Endocytosis
  • Humans
  • Inositol Phosphates / metabolism
  • Ligands
  • NF-kappa B / metabolism
  • Proteins / metabolism
  • Receptors, Chemokine / genetics
  • Receptors, Chemokine / metabolism*
  • Receptors, Chemokine / physiology*
  • Receptors, G-Protein-Coupled / metabolism
  • Receptors, G-Protein-Coupled / physiology*
  • Signal Transduction*
  • Type C Phospholipases / metabolism

Substances

  • Chemokines
  • Cyclic AMP Response Element-Binding Protein
  • Inositol Phosphates
  • Ligands
  • NF-kappa B
  • Proteins
  • Receptors, Chemokine
  • Receptors, G-Protein-Coupled
  • Type C Phospholipases