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

Jeffrey B. Woodruff, Beatriz Ferreira Gomes, Per O. Widlund, Julia Mahamid, Alf Honigmann, Anthony A. Hyman

Research output: Contribution to journalArticle

133 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)1066-1077.e10
JournalCell
Volume169
Issue number6
DOIs
Publication statusPublished - Jun 1 2017
Externally publishedYes

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Keywords

  • Centrosome
  • microtubule-organizing center
  • pericentriolar material
  • phase separation
  • Polo Kinase
  • scaffold
  • SPD-5
  • TPX2
  • XMAP215

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Woodruff, J. B., Ferreira Gomes, B., Widlund, P. O., Mahamid, J., Honigmann, A., & Hyman, A. A. (2017). The Centrosome Is a Selective Condensate that Nucleates Microtubules by Concentrating Tubulin. Cell, 169(6), 1066-1077.e10. https://doi.org/10.1016/j.cell.2017.05.028