A role for the vesicle tethering protein, p115, in the post-mitotic stacking of reassembling Golgi cisternae in a cell-free system.
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.
Shorter
J
J
Cell Biology Laboratory, Imperial Cancer Research Fund, London WC2A 3PX, United Kingdom. shorter@icrf.icnet.uk
Warren
G
G
D000251:Adenosine Triphosphatases; D000818:Animals; D001324:Autoantigens; D002352:Carrier Proteins / Q000032:analysis / Q000502:physiology*; D018797:Cell Cycle Proteins / Q000032:analysis / Q000502:physiology; D002499:Centrifugation, Density Gradient; D003600:Cytosol / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism; D005033:Ethylmaleimide / Q000494:pharmacology; D006056:Golgi Apparatus / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D000077614:Golgi Matrix Proteins; D006367:HeLa Cells; D006801:Humans; D007425:Intracellular Membranes / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D007700:Kinetics; D008099:Liver / Q000166:cytology / Q000378:metabolism; D008561:Membrane Fusion / Q000187:drug effects; D008565:Membrane Proteins / Q000032:analysis / Q000502:physiology*; D008938:Mitosis; D052066:N-Ethylmaleimide-Sensitive Proteins; D010446:Peptide Fragments / Q000502:physiology; D051381:Rats; D050600:SNARE Proteins; D012995:Solubility; D000074405:Valosin Containing Protein; D033921:Vesicular Transport Proteins
D001324:Autoantigens; D002352:Carrier Proteins; D018797:Cell Cycle Proteins; C108705:GORASP1 protein, human; D000077614:Golgi Matrix Proteins; C097741:Golgin subfamily A member 2; C491577:Gorasp1 protein, rat; D008565:Membrane Proteins; D010446:Peptide Fragments; D050600:SNARE Proteins; D033921:Vesicular Transport Proteins; C087574:macrogolgin; C091580:vesicular transport factor p115; D000251:Adenosine Triphosphatases; D052066:N-Ethylmaleimide-Sensitive Proteins; C498737:Nsf protein, rat; D000074405:Valosin Containing Protein; D005033:Ethylmaleimide