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The centrosome is a selective phase that nucleates microtubules by concentrating tubulin

View ORCID ProfileJeffrey B. Woodruff, Beatriz Ferreira Gomes, Per O. Widlund, Julia Mahamid, View ORCID ProfileAnthony A. Hyman
doi: https://doi.org/10.1101/093054
Jeffrey B. Woodruff
1Max Planck Institute of Molecular Cell Biology and Genetics, Pfotenhauerstrasse 108, 01307 Dresden, Germany
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  • For correspondence: hyman@mpi-cbg.de woodruff@mpi-cbg.de
Beatriz Ferreira Gomes
1Max Planck Institute of Molecular Cell Biology and Genetics, Pfotenhauerstrasse 108, 01307 Dresden, Germany
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Per O. Widlund
2Department of Chemistry and Molecular Biology, University of Gothenburg, Medicinaregatan 9 c, 40530 Gothenburg, Sweden
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Julia Mahamid
3Department of Molecular Structural Biology, Max Planck Institute of Biochemistry, Martinsried 82152, Germany
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Anthony A. Hyman
1Max Planck Institute of Molecular Cell Biology and Genetics, Pfotenhauerstrasse 108, 01307 Dresden, Germany
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  • For correspondence: hyman@mpi-cbg.de woodruff@mpi-cbg.de
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Abstract

Centrosomes are non-membrane-bound compartments that nucleate microtubule arrays. They consist of nanometer-scale centrioles surrounded by a micron-scale, dynamic assembly of protein called the pericentriolar material (PCM). To study how PCM forms a spherical compartment that nucleates microtubules, we reconstituted PCM-dependent microtubule nucleation in vitro using recombinant C.elegans proteins. We found that macromolecular crowding drives phase separation of the key PCM scaffold protein SPD-5 into spherical droplets that morphologically and dynamically resemble in vivo PCM. These SPD-5 droplets recruited the microtubule polymerase ZYG-9 (XMAP215 homologue) and the microtubule-stabilizing protein TPXL-1 (TPX2 homologue). Together, these three proteins concentrated tubulin ~4- fold over background, which was sufficient to reconstitute nucleation of microtubule asters in vitro. Our results suggest that in vivo PCM is a selective phase that organizes microtubule arrays through localized concentration of tubulin by microtubule effector proteins.

One Sentence Summary Phase separation of C. elegans centrosome proteins drive the formation of micron-sized microtubule organizing centers.

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Posted December 10, 2016.
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The centrosome is a selective phase that nucleates microtubules by concentrating tubulin
Jeffrey B. Woodruff, Beatriz Ferreira Gomes, Per O. Widlund, Julia Mahamid, Anthony A. Hyman
bioRxiv 093054; doi: https://doi.org/10.1101/093054
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The centrosome is a selective phase that nucleates microtubules by concentrating tubulin
Jeffrey B. Woodruff, Beatriz Ferreira Gomes, Per O. Widlund, Julia Mahamid, Anthony A. Hyman
bioRxiv 093054; doi: https://doi.org/10.1101/093054

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