Reconstitution of Golgi stacks from vesiculated Golgi membranes in permeabilized cells

Abstract

Golgi membranes are organized into stacks of flattened cisternae in the cytoplasm of eukaryotic cells. These membranes are constantly budding and receiving small transport vesicles during protein trafficking. In mammalian cells, during mitosis, the Golgi cisternae break down extensively into small vesicles and tubules, which are partitioned into the cytoplasm of the newly formed daughter cells, where they fuse and assemble into stacks of cisternae. These findings raise intriguing questions; how Golgi membranes maintain their organization in stacks of cisternae amidst the tremendous membrane flux in non-dividing cells and how is the organization of the Golgi regulated in association with the cell cycle events? The scope of this article is to describe an assay that reconstitutes the process of Golgi stack formation from vesiculated Golgi membranes (VGMs) in semi-intact cells and to highlight some of the recent findings that increase our understanding of the mechanism of Golgi stack assembly.