Yeast dynamin Vps1 associates with clathrin to facilitate vesicular trafficking and controls Golgi homeostasis

摘要

The yeast dynamin Vpsl acts cooperatively with many proteins at diverse cellular locations for endocytosis, protein sorting, and membrane fusion and fission. It has been proposed that Vpsl is functionally linked to clathrin heavy chain 1 (Chcl), but the question of how, where, and when they function together remains unknown. Here we report that Vpsl arrives at the Golgi after clathrin, and that loss of Vpsl leads to a shift in the cellular localization of clathrin to the late endosome and vacuole, not vice versa. Our two-hybrid-based approach provides evidence that full-length Vpsl and its truncated versions bind to the C-terminal region of the Chcl. Cells lacking both Vpsl and Chcl displayed more severe defects in carboxypeptidase Y (CPY) sorting at the Golgi than those in Vpsl-deficient cells. Further, these Vpsl fragments became dominant-negative for CPY sorting upon overexpression. These results suggest that Vpsl binds to Chcl and functions together at the Golgi for efficient Golgi-to-endosome membrane trafficking. In addition, we found that Vpsl, without the aid of clathrin, plays a role in controlling the number and turnover of late Golgi. (C) 2017 Elsevier GmbH. All rights reserved.