Abstract
The pericentriolar Golgi stacks are fragmented and found dispersed in mitotic mammalian cells. Addition of an antibody to the Golgi-associated protein GRASP65 inhibited Golgi fragmentation by mitotic cytosol in permeabilized cells. Microinjecting this antibody or the C-terminal fragment of GRASP65, which contains the antibody binding site, into normal rat kidney cells prevented entry into mitosis. Under these conditions the cells had completed S phase but were not in the prophase stage of mitosis. Fragmentation of the Golgi apparatus by nocodazole or Brefeldin A treatment prior to or post microinjection of the anti-GRASP65 antibody alleviated the block in mitotic entry. Based on our findings, we suggest that the pericentriolar Golgi organization is a sensor for controlling entry into mitosis in mammalian cells.
Publication types
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Comparative Study
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Amino Acid Sequence
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Animals
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Aphidicolin / pharmacology
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Binding Sites
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Biomarkers
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Brefeldin A / pharmacology
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Cells, Cultured
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Centrioles / metabolism
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Centrioles / physiology*
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Epitopes / metabolism
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Golgi Apparatus / drug effects
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Golgi Apparatus / metabolism
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Golgi Apparatus / physiology*
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Golgi Matrix Proteins
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Kidney / cytology
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Kidney / metabolism*
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Membrane Proteins / isolation & purification
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Membrane Proteins / metabolism
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Mitosis / drug effects
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Mitosis / physiology*
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Nocodazole / pharmacology
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Protein Structure, Tertiary
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Rats
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Recombinant Proteins / metabolism
Substances
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Biomarkers
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Epitopes
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Golgi Matrix Proteins
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Gorasp1 protein, rat
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Membrane Proteins
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Recombinant Proteins
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Brefeldin A
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Aphidicolin
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Nocodazole