Dynein and Mast/Orbit/CLASP have antagonistic roles in regulating kinetochore-microtubule plus-end dynamics
Dynein and Mast/Orbit/CLASP have antagonistic roles in regulating kinetochore-microtubule plus-end dynamics
Establishment and maintenance of the mitotic spindle requires the balanced activity of microtubule-associated proteins and motors. In this study we have addressed how the microtubule plus-end tracking protein Mast/Orbit/CLASP and cytoplasmic dynein regulate this process in Drosophila melanogaster embryos and S2 cells. We show that Mast accumulates at kinetochores early in mitosis, which is followed by a poleward streaming upon microtubule attachment. This leads to a reduction of Mast levels at kinetochores during metaphase and anaphase that depends largely on the microtubule minus end-directed motor cytoplasmic dynein. Surprisingly, we also found that co-depletion of Dynein rescues spindle bipolarity in Mast-depleted cells, while restoring normal microtubule poleward flux. Our results suggest that Mast and Dynein have antagonistic roles in the local regulation of microtubule plus-end dynamics at kinetochores, which are important for the maintenance of spindle bipolarity and normal spindle length.
Time Factors, Cytoplasmic Streaming, Dyneins, Mitosis, Microtubule dynamics, Transfection, Microtubules, Cell Line, Animals, Genetically Modified, Drosophila melanogaster, Larva, Plus-ends, Animals, Drosophila Proteins, Mast/Orbit/CLASP, Dynein, Kinetochore, Kinetochores, Microtubule-Associated Proteins, Signal Transduction
Time Factors, Cytoplasmic Streaming, Dyneins, Mitosis, Microtubule dynamics, Transfection, Microtubules, Cell Line, Animals, Genetically Modified, Drosophila melanogaster, Larva, Plus-ends, Animals, Drosophila Proteins, Mast/Orbit/CLASP, Dynein, Kinetochore, Kinetochores, Microtubule-Associated Proteins, Signal Transduction
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