Format

Send to

Choose Destination
J Cell Biol. 2017 Apr 3;216(4):983-997. doi: 10.1083/jcb.201607060. Epub 2017 Mar 6.

Structural analysis of the role of TPX2 in branching microtubule nucleation.

Author information

1
Department of Molecular Biology, Princeton University, Princeton, NJ 08544.
2
Department of Chemical and Biological Engineering, Princeton University, Princeton, NJ 08544.
3
Department of Molecular Biology, Princeton University, Princeton, NJ 08544 spetry@princeton.edu.

Abstract

The mitotic spindle consists of microtubules (MTs), which are nucleated by the γ-tubulin ring complex (γ-TuRC). How the γ-TuRC gets activated at the right time and location remains elusive. Recently, it was uncovered that MTs nucleate from preexisting MTs within the mitotic spindle, which requires the protein TPX2, but the mechanism basis for TPX2 action is unknown. Here, we investigate the role of TPX2 in branching MT nucleation. We establish the domain organization of Xenopus laevis TPX2 and define the minimal TPX2 version that stimulates branching MT nucleation, which we find is unrelated to TPX2's ability to nucleate MTs in vitro. Several domains of TPX2 contribute to its MT-binding and bundling activities. However, the property necessary for TPX2 to induce branching MT nucleation is contained within newly identified γ-TuRC nucleation activator motifs. Separation-of-function mutations leave the binding of TPX2 to γ-TuRC intact, whereas branching MT nucleation is abolished, suggesting that TPX2 may activate γ-TuRC to promote branching MT nucleation.

PMID:
28264915
PMCID:
PMC5379942
DOI:
10.1083/jcb.201607060
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for HighWire Icon for PubMed Central
Loading ...
Support Center