Epsin deficiency impairs endocytosis by stalling the actin-dependent invagination of endocytic clathrin-coated pits

Elife. 2014 Aug 13:3:e03311. doi: 10.7554/eLife.03311.

Abstract

Epsin is an evolutionarily conserved endocytic clathrin adaptor whose most critical function(s) in clathrin coat dynamics remain(s) elusive. To elucidate such function(s), we generated embryonic fibroblasts from conditional epsin triple KO mice. Triple KO cells displayed a dramatic cell division defect. Additionally, a robust impairment in clathrin-mediated endocytosis was observed, with an accumulation of early and U-shaped pits. This defect correlated with a perturbation of the coupling between the clathrin coat and the actin cytoskeleton, which we confirmed in a cell-free assay of endocytosis. Our results indicate that a key evolutionary conserved function of epsin, in addition to other roles that include, as we show here, a low affinity interaction with SNAREs, is to help generate the force that leads to invagination and then fission of clathrin-coated pits.

Keywords: Hip1R; SNARE; actin; clathrin-mediated endocytosis; cytokinesis; epsin.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actin Cytoskeleton / genetics
  • Actin Cytoskeleton / metabolism*
  • Actin Cytoskeleton / ultrastructure
  • Actins / genetics
  • Actins / metabolism*
  • Adaptor Proteins, Vesicular Transport / deficiency
  • Adaptor Proteins, Vesicular Transport / genetics*
  • Animals
  • Clathrin / genetics
  • Clathrin / metabolism*
  • Coated Pits, Cell-Membrane / genetics
  • Coated Pits, Cell-Membrane / metabolism*
  • Coated Pits, Cell-Membrane / ultrastructure
  • Embryo, Mammalian
  • Endocytosis / genetics*
  • Fibroblasts / metabolism
  • Fibroblasts / ultrastructure
  • Gene Expression
  • Mice
  • Mice, Knockout
  • Primary Cell Culture
  • Protein Binding
  • SNARE Proteins / genetics
  • SNARE Proteins / metabolism
  • Signal Transduction

Substances

  • Actins
  • Adaptor Proteins, Vesicular Transport
  • Clathrin
  • SNARE Proteins
  • epsin