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Published online 16 June 2003. doi:10.1083/jcb.200303048
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© The Rockefeller University Press, 0021-9525/2003/6/1067 $5.00
The Journal of Cell Biology, Volume 161, Number 6, 1067-1079

Article

 The localization and phosphorylation of p47 are important for Golgi disassembly–assembly during the cell cycle

Keiji Uchiyama1, Eija Jokitalo2, Mervi Lindman2, Mark Jackman3, Fumi Kano4, Masayuki Murata4, Xiaodong Zhang5 and Hisao Kondo1,6

1 Cambridge Institute for Medical Research, University of Cambridge, Cambridge CB2 2XY, UK
2 Institute of Biotechnology, Electron Microscopy Unit, University of Helsinki, FIN-00014 Helsinki, Finland
3 Wellcome/CRC Institute, University of Cambridge, Cambridge CB2 1QR, UK
4 National Institute for Physiological Sciences, Okazaki 444-8585, Japan
5 Centre for Structural Biology, Imperial College of Science, Technology and Medicine, London SW7 2AZ, UK
6 PRESTO and SORST, Japan Science and Technology Corporation, Saitama 332-0012, Japan

Address correspondence to Hisao Kondo, Cambridge Institute for Medical Research (Rm. 5.36), University of Cambridge, Wellcome Trust/MRC Bldg., Hills Rd., Cambridge CB2 2XY, UK. Tel.: 44-1223-762632. Fax: 44-1223-762640. E-mail: hk228{at}cam.ac.uk

In mammalian cells, the Golgi apparatus is disassembled at the onset of mitosis and reassembled at the end of mitosis. This disassembly–reassembly is generally believed to be essential for the equal partitioning of Golgi into two daughter cells. For Golgi disassembly, membrane fusion, which is mediated by NSF and p97, needs to be blocked. For the NSF pathway, the tethering of p115-GM130 is disrupted by the mitotic phosphorylation of GM130, resulting in the inhibition of NSF-mediated fusion. In contrast, the p97/p47 pathway does not require p115-GM130 tethering, and its mitotic inhibitory mechanism has been unclear. Now, we have found that p47, which mainly localizes to the nucleus during interphase, is phosphorylated on Serine-140 by Cdc2 at mitosis. The phosphorylated p47 does not bind to Golgi membranes. An in vitro assay shows that this phosphorylation is required for Golgi disassembly. Microinjection of p47(S140A), which is unable to be phosphorylated, allows the cell to keep Golgi stacks during mitosis and has no effect on the equal partitioning of Golgi into two daughter cells, suggesting that Golgi fragmentation-dispersion may not be obligatory for equal partitioning even in mammalian cells.

Key Words: Golgi; p47; p97; phosphorylation; mitosis

The online version of this article includes supplemental material.

* Abbreviations used in this paper: GalT, ß1,4-galactosyltransferase; Ser, serine; Thr, threonine.


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