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.
Cell Biology Laboratory, Imperial Cancer Research Fund, London WC2A 3PX, United Kingdom. shorter@icrf.icnet.uk
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Shorter
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Warren
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