Compartmentalized Toxoplasma EB1 bundles spindle microtubules to secure accurate chromosome segregation

Mol Biol Cell. 2015 Oct 14. pii: mbc.E15-06-0437. [Epub ahead of print]

 

Chen CT¹, Kelly M¹, de Leon J², Nwagbara B¹, Ebbert P¹, Ferguson DJ³, Lowery LA¹, Morrissette N⁴, Gubbels MJ⁵.

 

Toxoplasma gondii replicates asexually by a unique internal budding process characterized by interwoven closed mitosis and cytokinesis. Although it is known that the centrosome coordinates these processes, the spatiotemporal organization of mitosis remains poorly defined. Here we demonstrate that centrosome positioning around the nucleus may signal spindle assembly: spindle microtubules (MTs) are first assembled when the centrosome moves to the basal side, and become extensively acetylated after the duplicated centrosomes reposition to the apical side. We also tracked the spindle MTs using the MT plus-end binding protein TgEB1. Endowed by a C-terminal NLS, TgEB1 resides in the nucleoplasm in interphase and associates with the spindle MTs during mitosis. TgEB1 also associates with the subpellicular MTs at the growing end of daughter buds toward the completion of karyokinesis. Depletion of TgEB1 results in escalated disintegration of kinetochore clustering. Furthermore, we show that TgEB1's MT association in Toxoplasma and in a heterologous system (Xenopus) is based on the same principles. Finally, overexpression of a high MT affinity TgEB1 mutant promotes the formation of overstabilized MT bundles resulting in avulsion of otherwise tightly clustered kinetochores. Overall, we conclude that centrosome position controls spindle activity and that TgEB1 is critical for mitotic integrity.
© 2015 by The American Society for Cell Biology.

PMID:

26466679

[PubMed - as supplied by publisher]