A Role for the Vesicle Tethering Protein, p115, in the Post-mitotic Stacking of Reassembling Golgi Cisternae in a Cell-free System

doi:10.1083/jcb.146.1.57

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J. Cell Biol., Volume 146, Number 1, July 12, 1999 57-70
Copyright © 1999 by The Rockefeller University Press.

A Role for the Vesicle Tethering Protein, p115, in the Post-mitotic Stacking of Reassembling Golgi Cisternae in a Cell-free System

James Shorter^a and Graham Warren^a
^a Cell Biology Laboratory, Imperial Cancer Research Fund, London WC2A 3PX, United Kingdom

Correspondence to: James Shorter, Cell Biology Laboratory, Imperial Cancer Research Fund, 44 Lincoln's Inn Fields, London WC2A 3PX, UK., shorter{at}icrf.icnet.uk (E-mail), 0171-269-3403 (phone), 0171-269-3417 (fax)

During telophase, Golgi cisternae are regenerated and stackedfrom a heterogeneous population of tubulovesicular clusters.A cell-free system that reconstructs these events has revealedthat cisternal regrowth requires interplay between soluble factorsand soluble N-ethylmaleimide (NEM)–sensitive fusion protein(NSF) attachment protein receptors (SNAREs) via two intersectingpathways controlled by the ATPases, p97 and NSF. Golgi reassemblystacking protein 65 (GRASP65), an NEM-sensitive membrane-boundcomponent, is required for the stacking process. NSF-mediatedcisternal 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 withits receptors, in stacking p97 generated cisternae. Temporalanalysis suggests that p115 plays a transient role in stackingthat may be upstream of GRASP65-mediated stacking. These resultsimplicate p115 and its receptors in the initial alignment anddocking of single cisternae that may be an important prerequisitefor stack formation.

Key Words: Golgi apparatus, mitosis, p115, tethering, stack

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