Phosphorylation of the Vesicle-tethering Protein p115 by a Casein Kinase II-like Enzyme Is Required for Golgi Reassembly from Isolated Mitotic Fragments

doi:10.1083/jcb.150.3.475

A correction to this article has been published: J. Cell Biol. 150 (4) 937
Published online 7 August 2000. doi:10.1083/jcb.150.3.475

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© The Rockefeller University Press, 0021-9525/2000/8/475/ $5.00
The Journal of Cell Biology, Volume 150, Number 3, August 7, 2000 475-488

Original Article

Phosphorylation of the Vesicle-tethering Protein p115 by a Casein Kinase II–like Enzyme Is Required for Golgi Reassembly from Isolated Mitotic Fragments

A. Barbara Dirac-Svejstrup^a, James Shorter^c, M. Gerard Waters^b, and Graham Warren^c
^a Cell Biology Laboratory, Imperial Cancer Research Fund, London WC2A 3PX, United Kingdom
^b Department of Molecular Biology, Princeton University, Princeton, New Jersey 08544
^c Department of Cell Biology, Yale University School of Medicine, New Haven, Connecticut 06520-8002

Correspondence to: Graham Warren, Department of Cell Biology, SHM, C441, Yale University School of Medicine, 333 Cedar St., P.O. Box 208002, New Haven, CT 06520-8002. Tel:(203) 785-5058 Fax:(203) 785-4301 E-mail:graham.warren{at}yale.edu.

Coat protein I (COPI) transport vesicles can be tethered toGolgi membranes by a complex of fibrous, coiled-coil proteinscomprising p115, Giantin and GM130. p115 has been postulatedto act as a bridge, linking Giantin on the vesicle to GM130on the Golgi membrane. Here we show that the acidic COOH terminusof p115 mediates binding to both GM130 and Giantin as well aslinking the two together. Phosphorylation of serine 941 withinthis acidic domain enhances the binding as well as the linkbetween them. Phosphorylation is mediated by casein kinase II(CKII) or a CKII-like kinase. Surprisingly, the highly conservedNH₂-terminal head domain of p115 is not required for the NSF(N-ethylmaleimide–sensitive fusion protein)–catalyzedreassembly of cisternae from mitotic Golgi fragments in a cell-freesystem. However, the ability of p115 to link GM130 to Giantinand the phosphorylation of p115 at serine 941 are required forNSF-catalyzed cisternal regrowth. p115 phosphorylation may berequired for the transition from COPI vesicle tethering to COPIvesicle docking, an event that involves the formation of t-SNARE(trans–soluble NSF attachment protein [SNAP] receptor)complexes.

Key Words: p115, GM130, Giantin, CKII, tethering

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