Export from Pericentriolar Endocytic Recycling Compartment to Cell Surface Depends on Stable, Detyrosinated (Glu) Microtubules and Kinesin

Abstract

A significant fraction of internalized transferrin (Tf) concentrates in the endocytic recycling compartment (ERC), which is near the microtubule-organizing center in many cell types. Tf then recycles back to the cell surface. The mechanisms controlling the localization, morphology, and function of the ERC are not fully understood. We examined the relationship of Tf trafficking with microtubules (MTs), specifically the subset of stable, detyrosinated Glu MTs. We found some correlation between the level of stable Glu MTs and the distribution of the ERC; in cells with low levels of Glu MTs concentrated near to the centriole, the ERC was often tightly clustered, whereas in cells with higher levels of Glu MTs throughout the cell, the ERC was more dispersed. The clustered ERC in Chinese hamster ovary cells became dispersed when the level of Glu MTs was increased with taxol treatment. Furthermore, in a temperature-sensitive Chinese hamster ovary cell line (B104-5), the cells had more Glu MTs when the ERC became dispersed at elevated temperature. Microinjecting purified anti-Glu tubulin antibody into B104-5 cells at elevated temperature induced the redistribution of the ERC to a tight cluster. Microinjection of anti-Glu tubulin antibody slowed recycling of Tf to the cell surface without affecting Tf internalization or delivery to the ERC. Similar inhibition of Tf recycling was caused by microinjecting anti-kinesin antibody. These results suggest that stable Glu MTs and kinesin play a role in the organization of the ERC and in facilitating movement of vesicles from the ERC to the cell surface.

Regulation of endosomal membrane traffic by a Gadkin/AP-1/kinesin KIF5 complex

“…S5D). These data are consistent with a role for kinesin-dependent microtubular transport in regulating the dynamics of perinuclear endosomes (9). In agreement with this, depletion of the ubiquitous kinesin isoform KIF5B by two different siRNA oligonucleotides (Fig.…”

RalA-exocyst-dependent Recycling Endosome Trafficking Is Required for the Completion of Cytokinesis

“…3 However, the events upstream of this polarized vesicle delivery are not completely understood as both Sec8 and Sec5 seem to be mobilized in a RalA-independent manner. The classic Rappaport experiment (25) and additional recent studies indicated that the centrosome is able to influence polarized vesicle trafficking from the RE (20,26,27). We found that both RalA and the exocyst are associated with the centrosome, whereas disruption of their function causes defects in the late stage of cytokinesis, similar to defects resulting from a loss of centrosome function.…”

Kinesin 5B Is Necessary for Delivery of Membrane and Receptors during FcγR-Mediated Phagocytosis

“…We cannot be certain that phagocytosis initiates a global kinesin-dependent movement of recycling endosomes to the plasma membrane, but the reduced Rab11-EGFP on nascent phagosomes in kinesin-mutant cells suggest that localized trafficking of recycling endosomes may occur to active plasma membranes engaged in phagocytosis. Selective trafficking of recycling endosomes may occur on stabilized MT subsets, which have been shown to enhance kinesin-based transport (33). We have observed MTs penetrating into sites of phagocytosis, and these MTs may serve as tracks for kinesin-mediated recycling endosome delivery (11,12,34).…”