The Mitotic Phosphorylation Cycle of the cis-Golgi Matrix Protein GM130

doi:10.1083/jcb.149.2.341

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© The Rockefeller University Press, 0021-9525/2000/4/341/ $5.00
The Journal of Cell Biology, Volume 149, Number 2, April 17, 2000 341-356

Original Article

The Mitotic Phosphorylation Cycle of the cis-Golgi Matrix Protein GM130

Martin Lowe^a, Nicholas K. Gonatas^b, and Graham Warren^a
^a Cell Biology Laboratory, Imperial Cancer Research Fund, London WC2A 3PX, United Kingdom
^b Department of Pathology and Laboratory Medicine, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104

Correspondence to: Martin Lowe, School of Biological Sciences, University of Manchester, 2.205 Stopford Building, Oxford Road, Manchester M13 9PT, UK. Tel:0161-275-5387 Fax:0161-275-5082 E-mail:lowe{at}man.ac.uk.

The cis-Golgi matrix protein GM130 is phosphorylated in mitosison serine 25. Phosphorylation inhibits binding to p115, a vesicle-tetheringprotein, and has been implicated as an important step in themitotic Golgi fragmentation process. We have generated an antibodythat specifically recognizes GM130 phosphorylated on serine25, and used this antibody to study the temporal regulationof phosphorylation in vivo. GM130 is phosphorylated in prophaseas the Golgi complex starts to break down, and remains phosphorylatedduring further breakdown and partitioning of the Golgi fragmentsin metaphase and anaphase. In telophase, GM130 is dephosphorylatedas the Golgi fragments start to reassemble. The timing of phosphorylationand dephosphorylation correlates with the dissociation and reassociationof p115 with Golgi membranes. GM130 phosphorylation and p115dissociation appear specific to mitosis, since they are notinduced by several drugs that trigger nonmitotic Golgi fragmentation.The phosphatase responsible for dephosphorylation of mitoticGM130 was identified as PP2A. The active species was identifiedas heterotrimeric phosphatase containing the B ${alpha}$ regulatory subunit,suggesting a role for this isoform in the reassembly of mitoticGolgi membranes at the end of mitosis.

Key Words: Golgi, mitosis, GM130, phosphorylation, protein phosphatase2A

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