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J. Biol. Chem., Vol. 268, Issue 6, 4050-4054, 02, 1993
RA Stuart, D Mackay, J Adamczewski and G Warren
It has previously been shown that exocytic and endocytic membrane traffic
are inhibited in mitotic mammalian cells. Here we have used a cell-free
intra-Golgi transport assay supplemented with heterologous cytosols to
mimic this effect in vitro. Cytosols with high histone kinase activity,
made either from mitotic cells or by cyclin A treatment of interphase
cells, inhibited intra-Golgi transport by up to 75%. Inhibition of
transport was reversed by the kinase inhibitor staurosporine or by
reduction in ATP levels leading to inactivation of histone kinase. The data
indicate that cell cycle control of intra- Golgi transport is due to a
reversible modification of cytosol, and this assay system may be used to
study the molecular mechanism of mitotic transport inhibition in mammalian
cells.
Inhibition of intra-Golgi transport in vitro by mitotic kinase
Cell Biology Laboratory, Imperial Cancer Research Fund, London, United Kingdom.
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