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Regulation of Kif15 localization and motility by the C-terminus of TPX2 and microtubule dynamics

Mitotic motor proteins generate force to establish and maintain spindle bipolarity, but how they are temporally and spatially regulated in vivo is unclear. Prior work demonstrated that a microtubule-associated protein, TPX2, targets kinesin-5 and kinesin-12 motors to spindle microtubules. The C-term...

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Autores principales: Mann, Barbara J., Balchand, Sai K., Wadsworth, Patricia
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The American Society for Cell Biology 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5221630/
https://www.ncbi.nlm.nih.gov/pubmed/27852894
http://dx.doi.org/10.1091/mbc.E16-06-0476
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author Mann, Barbara J.
Balchand, Sai K.
Wadsworth, Patricia
author_facet Mann, Barbara J.
Balchand, Sai K.
Wadsworth, Patricia
author_sort Mann, Barbara J.
collection PubMed
description Mitotic motor proteins generate force to establish and maintain spindle bipolarity, but how they are temporally and spatially regulated in vivo is unclear. Prior work demonstrated that a microtubule-associated protein, TPX2, targets kinesin-5 and kinesin-12 motors to spindle microtubules. The C-terminal domain of TPX2 contributes to the localization and motility of the kinesin-5, Eg5, but it is not known whether this domain regulates kinesin-12, Kif15. We found that the C-terminal domain of TPX2 contributes to the localization of Kif15 to spindle microtubules in cells and suppresses motor walking in vitro. Kif15 and Eg5 are partially redundant motors, and overexpressed Kif15 can drive spindle formation in the absence of Eg5 activity. Kif15-dependent bipolar spindle formation in vivo requires the C-terminal domain of TPX2. In the spindle, fluorescent puncta of GFP-Kif15 move toward the equatorial region at a rate equivalent to microtubule growth. Reduction of microtubule growth with paclitaxel suppresses GFP-Kif15 motility, demonstrating that dynamic microtubules contribute to Kif15 behavior. Our results show that the C-terminal region of TPX2 regulates Kif15 in vitro, contributes to motor localization in cells, and is required for Kif15 force generation in vivo and further reveal that dynamic microtubules contribute to Kif15 behavior in vivo.
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spelling pubmed-52216302017-03-16 Regulation of Kif15 localization and motility by the C-terminus of TPX2 and microtubule dynamics Mann, Barbara J. Balchand, Sai K. Wadsworth, Patricia Mol Biol Cell Articles Mitotic motor proteins generate force to establish and maintain spindle bipolarity, but how they are temporally and spatially regulated in vivo is unclear. Prior work demonstrated that a microtubule-associated protein, TPX2, targets kinesin-5 and kinesin-12 motors to spindle microtubules. The C-terminal domain of TPX2 contributes to the localization and motility of the kinesin-5, Eg5, but it is not known whether this domain regulates kinesin-12, Kif15. We found that the C-terminal domain of TPX2 contributes to the localization of Kif15 to spindle microtubules in cells and suppresses motor walking in vitro. Kif15 and Eg5 are partially redundant motors, and overexpressed Kif15 can drive spindle formation in the absence of Eg5 activity. Kif15-dependent bipolar spindle formation in vivo requires the C-terminal domain of TPX2. In the spindle, fluorescent puncta of GFP-Kif15 move toward the equatorial region at a rate equivalent to microtubule growth. Reduction of microtubule growth with paclitaxel suppresses GFP-Kif15 motility, demonstrating that dynamic microtubules contribute to Kif15 behavior. Our results show that the C-terminal region of TPX2 regulates Kif15 in vitro, contributes to motor localization in cells, and is required for Kif15 force generation in vivo and further reveal that dynamic microtubules contribute to Kif15 behavior in vivo. The American Society for Cell Biology 2017-01-01 /pmc/articles/PMC5221630/ /pubmed/27852894 http://dx.doi.org/10.1091/mbc.E16-06-0476 Text en © 2017 Mann, Balchand, and Wadsworth. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0). “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology.
spellingShingle Articles
Mann, Barbara J.
Balchand, Sai K.
Wadsworth, Patricia
Regulation of Kif15 localization and motility by the C-terminus of TPX2 and microtubule dynamics
title Regulation of Kif15 localization and motility by the C-terminus of TPX2 and microtubule dynamics
title_full Regulation of Kif15 localization and motility by the C-terminus of TPX2 and microtubule dynamics
title_fullStr Regulation of Kif15 localization and motility by the C-terminus of TPX2 and microtubule dynamics
title_full_unstemmed Regulation of Kif15 localization and motility by the C-terminus of TPX2 and microtubule dynamics
title_short Regulation of Kif15 localization and motility by the C-terminus of TPX2 and microtubule dynamics
title_sort regulation of kif15 localization and motility by the c-terminus of tpx2 and microtubule dynamics
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5221630/
https://www.ncbi.nlm.nih.gov/pubmed/27852894
http://dx.doi.org/10.1091/mbc.E16-06-0476
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