CARTS biogenesis requires VAP-lipid transfer protein complexes functioning at the endoplasmic reticulum-Golgi interface

Mol Biol Cell. 2015 Dec 15;26(25):4686-99. doi: 10.1091/mbc.E15-08-0599. Epub 2015 Oct 21.

Abstract

Vesicle-associated membrane protein-associated protein (VAP) is an endoplasmic reticulum (ER)-resident integral membrane protein that controls a nonvesicular mode of ceramide and cholesterol transfer from the ER to the Golgi complex by interacting with ceramide transfer protein and oxysterol-binding protein (OSBP), respectively. We report that VAP and its interacting proteins are required for the processing and secretion of pancreatic adenocarcinoma up-regulated factor, whose transport from the trans-Golgi network (TGN) to the cell surface is mediated by transport carriers called "carriers of the trans-Golgi network to the cell surface" (CARTS). In VAP-depleted cells, diacylglycerol level at the TGN was decreased and CARTS formation was impaired. We found that VAP forms a complex with not only OSBP but also Sac1 phosphoinositide phosphatase at specialized ER subdomains that are closely apposed to the trans-Golgi/TGN, most likely reflecting membrane contact sites. Immobilization of ER-Golgi contacts dramatically reduced CARTS production, indicating that association-dissociation dynamics of the two membranes are important. On the basis of these findings, we propose that the ER-Golgi contacts play a pivotal role in lipid metabolism to control the biogenesis of transport carriers from the TGN.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Ceramides / metabolism
  • Cholesterol / metabolism
  • Endoplasmic Reticulum / metabolism*
  • HEK293 Cells
  • Humans
  • Intercellular Signaling Peptides and Proteins
  • Lectins / genetics
  • Lectins / metabolism
  • Membrane Fusion / genetics
  • Protein Kinase C / genetics
  • Protein Kinase C / metabolism
  • Protein Serine-Threonine Kinases / genetics
  • Protein Serine-Threonine Kinases / metabolism
  • R-SNARE Proteins / genetics
  • R-SNARE Proteins / metabolism*
  • Vesicular Transport Proteins / genetics
  • Vesicular Transport Proteins / metabolism*
  • trans-Golgi Network / genetics
  • trans-Golgi Network / metabolism*

Substances

  • Ceramides
  • Intercellular Signaling Peptides and Proteins
  • Lectins
  • R-SNARE Proteins
  • VAPA protein, human
  • Vesicular Transport Proteins
  • ZG16B protein, human
  • Cholesterol
  • CERT1 protein, human
  • protein kinase D
  • Protein Serine-Threonine Kinases
  • Protein Kinase C