Rapid microtubule-independent dynamics of Cdc20 at kinetochores and centrosomes in mammalian cells.
Cdc20 is a substrate adaptor and activator of the anaphase-promoting complex/cyclosome (APC/C), the E3 ubiquitin ligase whose activity is required for anaphase onset and exit from mitosis. A green fluorescent protein derivative, Cdc20-GFP, bound to centrosomes throughout the cell cycle and to kinetochores from late prophase to late telophase. We ... mapped distinct domains of Cdc20 that are required for association with kinetochores and centrosomes. FRAP measurements revealed extremely rapid dynamics at the kinetochores (t1/2 = 5.1 s) and spindle poles (t1/2 = 4.7 s). This rapid turnover is independent of microtubules. Rapid transit of Cdc20 through kinetochores may ensure that spindle checkpoint signaling at unattached/relaxed kinetochores can continuously inhibit APC/CCdc20 targeting of anaphase inhibitors (securins) throughout the cell until all the chromosomes are properly attached to the mitotic spindle.
Mesh Terms:
Animals, Calcium-Binding Proteins, Cell Cycle Proteins, Centrosome, Fluorescence Recovery After Photobleaching, Hela Cells, Humans, Kinetochores, LLC-PK1 Cells, Ligases, Microscopy, Fluorescence, Microtubules, Mitosis, Models, Biological, Protein Kinases, Protein Structure, Tertiary, Protein-Serine-Threonine Kinases, Repressor Proteins, Saccharomyces cerevisiae Proteins, Time Factors, Ubiquitin-Protein Ligase Complexes
Animals, Calcium-Binding Proteins, Cell Cycle Proteins, Centrosome, Fluorescence Recovery After Photobleaching, Hela Cells, Humans, Kinetochores, LLC-PK1 Cells, Ligases, Microscopy, Fluorescence, Microtubules, Mitosis, Models, Biological, Protein Kinases, Protein Structure, Tertiary, Protein-Serine-Threonine Kinases, Repressor Proteins, Saccharomyces cerevisiae Proteins, Time Factors, Ubiquitin-Protein Ligase Complexes
J. Cell Biol.
Date: Sep. 02, 2002
PubMed ID: 12196507
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