A novel GTP-binding protein-adaptor protein complex responsible for export of Vangl2 from the trans Golgi network

Elife. 2013 Jan 8:2:e00160. doi: 10.7554/eLife.00160.

Abstract

Planar cell polarity (PCP) requires the asymmetric sorting of distinct signaling receptors to distal and proximal surfaces of polarized epithelial cells. We have examined the transport of one PCP signaling protein, Vangl2, from the trans Golgi network (TGN) in mammalian cells. Using siRNA knockdown experiments, we find that the GTP-binding protein, Arfrp1, and the clathrin adaptor complex 1 (AP-1) are required for Vangl2 transport from the TGN. In contrast, TGN export of Frizzled 6, which localizes to the opposing epithelial surface from Vangl2, does not depend on Arfrp1 or AP-1. Mutagenesis studies identified a YYXXF sorting signal in the C-terminal cytosolic domain of Vangl2 that is required for Vangl2 traffic and interaction with the μ subunit of AP-1. We propose that Arfrp1 exposes a binding site on AP-1 that recognizes the Vangl2 sorting motif for capture into a transport vesicle destined for the proximal surface of a polarized epithelial cell.DOI:http://dx.doi.org/10.7554/eLife.00160.001.

Keywords: Arf proteins; Human; TGN sorting; Vesicle coat proteins.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ADP-Ribosylation Factors / genetics
  • ADP-Ribosylation Factors / metabolism*
  • Adaptor Protein Complex 1 / metabolism
  • Adaptor Protein Complex gamma Subunits / metabolism
  • Adaptor Protein Complex mu Subunits / metabolism
  • Amino Acid Sequence
  • Animals
  • Binding Sites
  • COS Cells
  • Cadherins / metabolism
  • Cell Adhesion Molecules / metabolism
  • Cell Polarity*
  • Chlorocebus aethiops
  • Epithelial Cells / metabolism*
  • Frizzled Receptors / metabolism
  • HeLa Cells
  • Humans
  • Intracellular Signaling Peptides and Proteins / genetics
  • Intracellular Signaling Peptides and Proteins / metabolism*
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism*
  • Molecular Sequence Data
  • Mutation
  • Phosphorylation
  • Protein Binding
  • Protein Interaction Domains and Motifs
  • Protein Kinase C / metabolism
  • Protein Transport
  • RNA Interference
  • Receptor Protein-Tyrosine Kinases / metabolism
  • Transfection
  • trans-Golgi Network / metabolism*

Substances

  • Adaptor Protein Complex 1
  • Adaptor Protein Complex gamma Subunits
  • Adaptor Protein Complex mu Subunits
  • CELSR1 cadherin, human
  • Cadherins
  • Cell Adhesion Molecules
  • FZD6 protein, human
  • Frizzled Receptors
  • Intracellular Signaling Peptides and Proteins
  • Membrane Proteins
  • VANGL2 protein, human
  • protein kinase D
  • PTK7 protein, human
  • Receptor Protein-Tyrosine Kinases
  • Protein Kinase C
  • ADP-Ribosylation Factors
  • ARFRP1 protein, human

Grants and funding

The funder had no role in study design, data collection and interpretation, or the decision to submit the work for publication.