J. Cell Biol.
© The Rockefeller University Press
0021-9525/97/06/1211/18 $2.00
Volume 137, Number 6, June 16, 1997 1211-1228

Partitioning of the Golgi Apparatus during Mitosis in Living HeLa Cells

David T. Shima,^* Kasturi Haldar, Rainer Pepperkok,^§ Rose Watson,^* and Graham Warren^*

^* Cell Biology Laboratory, Imperial Cancer Research Fund, London WC2A, 3PX, UK;  Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, California 94305-5402; and ^§ Departement de Biologie Cellulaire, Universite de Geneve, CH-1211 Geneve 4, Switzerland

The Golgi apparatus of HeLa cells was fluorescently tagged with a green fluorescent protein (GFP), localized by attachment to the NH₂-terminal retention signal of N-acetylglucosaminyltransferase I (NAGT I). The location was confirmed by immunogold and immunofluorescence microscopy using a variety of Golgi markers. The behavior of the fluorescent Golgi marker was observed in fixed and living mitotic cells using confocal microscopy. By metaphase, cells contained a constant number of Golgi fragments dispersed throughout the cytoplasm. Conventional and cryoimmunoelectron microscopy showed that the NAGT I-GFP chimera (NAGFP)-positive fragments were tubulo-vesicular mitotic Golgi clusters. Mitotic conversion of Golgi stacks into mitotic clusters had surprisingly little effect on the polarity of Golgi membrane markers at the level of fluorescence microscopy. In living cells, there was little self-directed movement of the clusters in the period from metaphase to early telophase. In late telophase, the Golgi ribbon began to be reformed by a dynamic process of congregation and tubulation of the newly inherited Golgi fragments. The accuracy of partitioning the NAGFP-tagged Golgi was found to exceed that expected for a stochastic partitioning process. The results provide direct evidence for mitotic clusters as the unit of partitioning and suggest that precise regulation of the number, position, and compartmentation of mitotic membranes is a critical feature for the ordered inheritance of the Golgi apparatus.

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