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. 2017 Jun 1;169(6):1066-1077.e10.
doi: 10.1016/j.cell.2017.05.028.

The Centrosome Is a Selective Condensate that Nucleates Microtubules by Concentrating Tubulin

Jeffrey B Woodruff  1 , Beatriz Ferreira Gomes  2 , Per O Widlund  3 , Julia Mahamid  4 , Alf Honigmann  2 , Anthony A Hyman  5
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Free article

The Centrosome Is a Selective Condensate that Nucleates Microtubules by Concentrating Tubulin

Jeffrey B Woodruff et al. Cell. .
Free article

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 assembly of the key PCM scaffold protein SPD-5 into spherical condensates that morphologically and dynamically resemble in vivo PCM. These SPD-5 condensates recruited the microtubule polymerase ZYG-9 (XMAP215 homolog) and the microtubule-stabilizing protein TPXL-1 (TPX2 homolog). 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.

Keywords: Centrosome; Polo Kinase; SPD-5; TPX2; XMAP215; microtubule-organizing center; pericentriolar material; phase separation; scaffold.

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