Erv41p and Erv46p: new components of COPII vesicles involved in transport between the ER and Golgi complex

J Cell Biol. 2001 Feb 5;152(3):503-18. doi: 10.1083/jcb.152.3.503.

Abstract

Proteins contained on purified COPII vesicles were analyzed by matrix-assisted laser desorption ionization mass spectrometry combined with database searching. We identified four known vesicle proteins (Erv14p, Bet1p, Emp24p, and Erv25p) and an additional nine species (Yip3p, Rer1p, Erp1p, Erp2p, Erv29p, Yif1p, Erv41p, Erv46p, and Emp47p) that had not been localized to ER vesicles. Using antibodies, we demonstrate that these proteins are selectively and efficiently packaged into COPII vesicles. Three of the newly identified vesicle proteins (Erv29p, Erv41p, and Erv46p) represent uncharacterized integral membrane proteins that are conserved across species. Erv41p and Erv46p were further characterized. These proteins colocalized to ER and Golgi membranes and exist in a detergent-soluble complex that was isolated by immunoprecipitation. Yeast strains lacking Erv41p and/or Erv46p are viable but display cold sensitivity. The expression levels of Erv41p and Erv46p are interdependent such that Erv46p was reduced in an erv41Delta strain, and Erv41p was not detected in an erv46Delta strain. When the erv41Delta or ev46Delta alleles were combined with other mutations in the early secretory pathway, altered growth phenotypes were observed in some of the double mutant strains. A cell-free assay that reproduces transport between the ER and Golgi indicates that deletion of the Erv41p-Erv46p complex influences the membrane fusion stage of transport.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • COP-Coated Vesicles / chemistry
  • COP-Coated Vesicles / metabolism*
  • Cell Fractionation
  • Databases, Factual
  • Endoplasmic Reticulum / chemistry
  • Endoplasmic Reticulum / metabolism*
  • Fungal Proteins / genetics
  • Fungal Proteins / metabolism*
  • Golgi Apparatus / chemistry
  • Golgi Apparatus / metabolism*
  • Immunoblotting
  • Intracellular Membranes / metabolism*
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism*
  • Plasmids / genetics
  • Plasmids / metabolism
  • Precipitin Tests
  • Protein Transport / physiology*
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Yeasts / genetics
  • Yeasts / metabolism

Substances

  • Fungal Proteins
  • Membrane Proteins
  • Recombinant Fusion Proteins