A role for the vesicle tethering protein, p115, in the post-mitotic stacking of reassembling Golgi cisternae in a cell-free system

J Cell Biol. 1999 Jul 12;146(1):57-70. doi: 10.1083/jcb.146.1.57.

Abstract

During telophase, Golgi cisternae are regenerated and stacked from a heterogeneous population of tubulovesicular clusters. A cell-free system that reconstructs these events has revealed that cisternal regrowth requires interplay between soluble factors and soluble N-ethylmaleimide (NEM)-sensitive fusion protein (NSF) attachment protein receptors (SNAREs) via two intersecting pathways controlled by the ATPases, p97 and NSF. Golgi reassembly stacking protein 65 (GRASP65), an NEM-sensitive membrane-bound component, is required for the stacking process. NSF-mediated cisternal regrowth requires a vesicle tethering protein, p115, which we now show operates through its two Golgi receptors, GM130 and giantin. p97-mediated cisternal regrowth is p115-independent, but we now demonstrate a role for p115, in conjunction with its receptors, in stacking p97 generated cisternae. Temporal analysis suggests that p115 plays a transient role in stacking that may be upstream of GRASP65-mediated stacking. These results implicate p115 and its receptors in the initial alignment and docking of single cisternae that may be an important prerequisite for stack formation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphatases
  • Animals
  • Autoantigens
  • Carrier Proteins / analysis
  • Carrier Proteins / physiology*
  • Cell Cycle Proteins / analysis
  • Cell Cycle Proteins / physiology
  • Centrifugation, Density Gradient
  • Cytosol / chemistry
  • Cytosol / drug effects
  • Cytosol / metabolism
  • Ethylmaleimide / pharmacology
  • Golgi Apparatus / drug effects
  • Golgi Apparatus / metabolism*
  • Golgi Apparatus / ultrastructure
  • Golgi Matrix Proteins
  • HeLa Cells
  • Humans
  • Intracellular Membranes / drug effects
  • Intracellular Membranes / metabolism*
  • Intracellular Membranes / ultrastructure
  • Kinetics
  • Liver / cytology
  • Liver / metabolism
  • Membrane Fusion / drug effects
  • Membrane Proteins / analysis
  • Membrane Proteins / physiology*
  • Mitosis*
  • N-Ethylmaleimide-Sensitive Proteins
  • Peptide Fragments / physiology
  • Rats
  • SNARE Proteins
  • Solubility
  • Valosin Containing Protein
  • Vesicular Transport Proteins*

Substances

  • Autoantigens
  • Carrier Proteins
  • Cell Cycle Proteins
  • GORASP1 protein, human
  • Golgi Matrix Proteins
  • Golgin subfamily A member 2
  • Gorasp1 protein, rat
  • Membrane Proteins
  • Peptide Fragments
  • SNARE Proteins
  • Vesicular Transport Proteins
  • macrogolgin
  • vesicular transport factor p115
  • Adenosine Triphosphatases
  • N-Ethylmaleimide-Sensitive Proteins
  • Nsf protein, rat
  • Valosin Containing Protein
  • Ethylmaleimide