BioID Performed on Golgi Enriched Fractions Identify C10orf76 as a GBF1 Binding Protein Essential for Golgi Maintenance and Secretion

Mol Cell Proteomics. 2019 Nov;18(11):2285-2297. doi: 10.1074/mcp.RA119.001645. Epub 2019 Sep 13.

Abstract

The Golgi-specific Brefeldin-A resistance factor 1 (GBF1) is the only large GEF that regulates Arf activation at the cis-Golgi and is actively recruited to membranes on an increase in Arf-GDP. Recent studies have revealed that GBF1 recruitment requires one or more heat-labile and protease-sensitive protein factor(s) (Quilty et al., 2018, J. Cell Science, 132). Proximity-dependent biotinylation (BioID) and mass spectrometry from enriched Golgi fractions identified GBF1 proximal proteins that may regulate its recruitment. Knockdown studies revealed C10orf76 to be involved in Golgi maintenance. We find that C10orf76 interacts with GBF1 and rapidly cycles on and off GBF1-positive Golgi structures. More importantly, its depletion causes Golgi fragmentation, alters GBF1 recruitment, and impairs secretion. Homologs were identified in most species, suggesting its presence in the last eukaryotic common ancestor.

Keywords: BioID; Cell biology; GBF1; GTPase; biotin; cell secretion; cellular organelles; golgi; imaging; membranes; secretome.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biotinylation
  • Carrier Proteins / metabolism*
  • Golgi Apparatus / metabolism*
  • Guanine Nucleotide Exchange Factors / metabolism*
  • HeLa Cells
  • Humans
  • Intracellular Membranes / metabolism*
  • Protein Binding
  • Protein Interaction Domains and Motifs
  • Protein Transport

Substances

  • Carrier Proteins
  • GBF1 protein, human
  • Guanine Nucleotide Exchange Factors

Grants and funding