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Elife. 2019 May 8;8. pii: e43890. doi: 10.7554/eLife.43890.

Spatiotemporal organization of branched microtubule networks.

Author information

1
Department of Chemical and Biological Engineering, Princeton University, Princeton, United States.
2
Department of Mechanical and Aerospace Engineering, Princeton University, Princeton, United States.
3
Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, United States.
4
Department of Physics, Princeton University, Princeton, United States.
5
Department of Molecular Biology, Princeton University, Princeton, United States.

Abstract

To understand how chromosomes are segregated, it is necessary to explain the precise spatiotemporal organization of microtubules (MTs) in the mitotic spindle. We use Xenopus egg extracts to study the nucleation and dynamics of MTs in branched networks, a process that is critical for spindle assembly. Surprisingly, new branched MTs preferentially originate near the minus-ends of pre-existing MTs. A sequential reaction model, consisting of deposition of nucleation sites on an existing MT, followed by rate-limiting nucleation of branches, reproduces the measured spatial profile of nucleation, the distribution of MT plus-ends and tubulin intensity. By regulating the availability of the branching effectors TPX2, augmin and γ-TuRC, combined with single-molecule observations, we show that first TPX2 is deposited on pre-existing MTs, followed by binding of augmin/γ-TuRC to result in the nucleation of branched MTs. In sum, regulating the localization and kinetics of nucleation effectors governs the architecture of branched MT networks.

KEYWORDS:

TPX2; Xenopus; augmin; branching microtubule nucleation; cell biology; microtubules; mitotic spindle; physics of living systems; xenopus

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