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Liquid-liquid phase separation of the chromosomal passenger complex drives parallel bundling of midzone microtubules

View ORCID ProfileEwa Niedzialkowska, Tan M. Truong, Luke A. Eldredge, Stefanie Redemann, Denis Chretien, View ORCID ProfileP. Todd Stukenberg
doi: https://doi.org/10.1101/2022.01.07.475460
Ewa Niedzialkowska
1Department of Biochemistry and Molecular Biology, University of Virginia, School of Medicine, Charlottesville, VA, USA
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  • ORCID record for Ewa Niedzialkowska
Tan M. Truong
2Department of Cell Biology, University of Virginia, School of Medicine, Charlottesville, VA, USA
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Luke A. Eldredge
2Department of Cell Biology, University of Virginia, School of Medicine, Charlottesville, VA, USA
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Stefanie Redemann
1Department of Biochemistry and Molecular Biology, University of Virginia, School of Medicine, Charlottesville, VA, USA
3Department of Molecular Physiology and Biological Physics, University of Virginia, School of Medicine, Charlottesville, VA, USA
4Center for Membrane and Cell Physiology, University of Virginia, School of Medicine, Charlottesville, VA, USA
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Denis Chretien
5Univ Rennes, CNRS, IGDR (Institut de Génétique et Développement de Rennes) - UMR 6290, F-35000 Rennes, France
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P. Todd Stukenberg
1Department of Biochemistry and Molecular Biology, University of Virginia, School of Medicine, Charlottesville, VA, USA
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  • For correspondence: pts7h@virginia.edu
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Abstract

The spindle midzone is a dynamic structure that forms during anaphase, mediates chromosome segregation, and provides a signaling platform to position the cleavage furrow. The spindle midzone comprises two antiparallel bundles of microtubules (MTs) but the process of their formation is poorly understood. Here, we show that the Chromosomal Passenger Complex (CPC) undergoes liquid-liquid phase separation (LLPS) to generate parallel MT bundles in vitro when incubated with free tubulin and GTP. MT bundles emerge from CPC droplets with protruding minus-ends that then grow into long, tapered MT structures. During this growth, the CPC in condensates apparently reorganize to coat and bundle the resulting MT structures. CPC mutants attenuated for LLPS or MT binding prevented the generation of parallel MT bundles in vitro and reduced the number of MTs present at spindle midzones in HeLa cells. Our data uncovers a kinase-independent function of the CPC and provides models for how cells generate parallel-bundled MT structures that are important for the assembly of the mitotic spindle.

Competing Interest Statement

The authors have declared no competing interest.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
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Posted January 09, 2022.
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Liquid-liquid phase separation of the chromosomal passenger complex drives parallel bundling of midzone microtubules
Ewa Niedzialkowska, Tan M. Truong, Luke A. Eldredge, Stefanie Redemann, Denis Chretien, P. Todd Stukenberg
bioRxiv 2022.01.07.475460; doi: https://doi.org/10.1101/2022.01.07.475460
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Liquid-liquid phase separation of the chromosomal passenger complex drives parallel bundling of midzone microtubules
Ewa Niedzialkowska, Tan M. Truong, Luke A. Eldredge, Stefanie Redemann, Denis Chretien, P. Todd Stukenberg
bioRxiv 2022.01.07.475460; doi: https://doi.org/10.1101/2022.01.07.475460

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