Dynamin-dependent transferrin receptor recycling by endosome-derived clathrin-coated vesicles

Mol Biol Cell. 2002 Jan;13(1):169-82. doi: 10.1091/mbc.01-07-0380.

Abstract

Previously we described clathrin-coated buds on tubular early endosomes that are distinct from those at the plasma membrane and the trans-Golgi network. Here we show that these clathrin-coated buds, like plasma membrane clathrin-coated pits, contain endogenous dynamin-2. To study the itinerary that is served by endosome-derived clathrin-coated vesicles, we used cells that overexpressed a temperature-sensitive mutant of dynamin-1 (dynamin-1(G273D)) or, as a control, dynamin-1 wild type. In dynamin-1(G273D)-expressing cells, 29-36% of endocytosed transferrin failed to recycle at the nonpermissive temperature and remained associated with tubular recycling endosomes. Sorting of endocytosed transferrin from fluid-phase endocytosed markers in early endosome antigen 1-labeled sorting endosomes was not inhibited. Dynamin-1(G273D) associated with accumulated clathrin-coated buds on extended tubular recycling endosomes. Brefeldin A interfered with the assembly of clathrin coats on endosomes and reduced the extent of transferrin recycling in control cells but did not further affect recycling by dynamin-1(G273D)-expressing cells. Together, these data indicate that the pathway from recycling endosomes to the plasma membrane is mediated, at least in part, by endosome-derived clathrin-coated vesicles in a dynamin-dependent manner.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blotting, Western
  • Brefeldin A / pharmacology
  • Clathrin-Coated Vesicles / drug effects
  • Clathrin-Coated Vesicles / metabolism*
  • Clathrin-Coated Vesicles / ultrastructure
  • Dynamin I
  • Dynamins
  • Endocytosis / physiology
  • Endosomes / drug effects
  • Endosomes / metabolism*
  • Endosomes / ultrastructure
  • GTP Phosphohydrolases / immunology
  • GTP Phosphohydrolases / metabolism*
  • GTP Phosphohydrolases / ultrastructure
  • HeLa Cells
  • Hot Temperature
  • Humans
  • Microscopy, Confocal
  • Microscopy, Immunoelectron
  • Mutation
  • Protein Synthesis Inhibitors / pharmacology
  • Protein Transport
  • Receptors, Transferrin / metabolism*
  • Transferrin / metabolism
  • Tumor Cells, Cultured

Substances

  • Protein Synthesis Inhibitors
  • Receptors, Transferrin
  • Transferrin
  • Brefeldin A
  • Dynamin I
  • GTP Phosphohydrolases
  • Dynamins