Oligomeric complexes link Rab5 effectors with NSF and drive membrane fusion via interactions between EEA1 and syntaxin 13

Cell. 1999 Aug 6;98(3):377-86. doi: 10.1016/s0092-8674(00)81966-2.

Abstract

SNAREs and Rab GTPases cooperate in vesicle transport through a mechanism yet poorly understood. We now demonstrate that the Rab5 effectors EEA1 and Rabaptin-5/Rabex-5 exist on the membrane in high molecular weight oligomers, which also contain NSF. Oligomeric assembly is modulated by the ATPase activity of NSF. Syntaxin 13, the t-SNARE required for endosome fusion, is transiently incorporated into the large oligomers via direct interactions with EEA1. This interaction is required to drive fusion, since both dominant-negative EEA1 and synthetic peptides encoding the FYVE Zn2+ finger hinder the interaction and block fusion. We propose a novel mechanism whereby oligomeric EEA1 and NSF mediate the local activation of syntaxin 13 upon membrane tethering and, by analogy with viral fusion proteins, coordinate the assembly of a fusion pore.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphatases / metabolism
  • Amino Acid Sequence
  • Autoantigens / metabolism
  • Biosensing Techniques
  • Carrier Proteins / metabolism*
  • Endosomes / drug effects
  • Endosomes / physiology
  • GTP Phosphohydrolases / metabolism
  • GTP-Binding Proteins / metabolism*
  • HeLa Cells
  • Humans
  • Intracellular Membranes / drug effects
  • Intracellular Membranes / physiology
  • Intracellular Membranes / ultrastructure
  • Membrane Fusion / drug effects
  • Membrane Fusion / physiology*
  • Membrane Proteins / metabolism*
  • Models, Biological
  • Molecular Sequence Data
  • N-Ethylmaleimide-Sensitive Proteins
  • Oligopeptides / chemistry
  • Oligopeptides / pharmacology
  • Peptide Fragments / chemistry
  • Peptide Fragments / pharmacology
  • Qa-SNARE Proteins
  • SNARE Proteins
  • Vesicular Transport Proteins*
  • Zinc Fingers
  • rab5 GTP-Binding Proteins

Substances

  • Autoantigens
  • Carrier Proteins
  • Membrane Proteins
  • Oligopeptides
  • Peptide Fragments
  • Qa-SNARE Proteins
  • SNARE Proteins
  • Vesicular Transport Proteins
  • early endosome antigen 1
  • Adenosine Triphosphatases
  • GTP Phosphohydrolases
  • GTP-Binding Proteins
  • N-Ethylmaleimide-Sensitive Proteins
  • rab5 GTP-Binding Proteins