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

EM van Dam, W Stoorvogel - Molecular biology of the cell, 2002 - Am Soc Cell Biol
EM van Dam, W Stoorvogel
Molecular biology of the cell, 2002Am Soc Cell Biol
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-1G273D) or, as a control, dynamin-1 wild type. In dynamin-1G273D–expressing …
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-1G273D) or, as a control, dynamin-1 wild type. In dynamin-1G273D–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-1G273D 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-1G273D–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.
Am Soc Cell Biol