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Published June 26, 2007 | Published + Supplemental Material
Journal Article Open

Recruitment of Polo Kinase to the Spindle Midzone during Cytokinesis Requires the Feo/Klp3A Complex

Abstract

Background: Polo-like kinases control multiple events during cell division, including mitotic entry, centrosome organization, spindle formation, chromosome segregation and cytokinesis. Their roles during cytokinesis, however, are not well understood because the requirement of these kinases during early stages of mitosis complicates the study of their functions after anaphase onset. Methodology/Principal Findings: We used time-lapse microscopy to analyze the dynamics of Polo::GFP in Drosophila tissue culture cells during mitosis. After anaphase onset, Polo::GFP concentrated at the spindle midzone, but also diffused along the entire length of the central spindle. Using RNA interference we demonstrate that the microtubule-associated proteins Feo and Klp3A are required for Polo recruitment to the spindle midzone, but not the kinesin Pavarotti as previously thought. Moreover, we show that Feo and Klp3A form a complex and that Polo co-localizes with both proteins during cytokinesis. Conclusion/Significance: Our results reveal that the Feo/Klp3A complex is necessary for Polo recruitment to the spindle midzone. A similar finding has also been recently reported in mammalian cells [1], suggesting that this basic mechanism has been conserved during evolution, albeit with some differences. Finally, since cleavage furrow formation and ingression are unaffected following feo RNAi, our data imply that Polo recruitment to the central spindle is not required for furrowing, but some other aspect of cytokinesis.

Additional Information

© 2007 D'Avino et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Received: March 27, 2007; Accepted: June 2, 2007; Published: June 27, 2007. This work was supported by a Cancer Research-UK Programme Grant (to DMG) and a BBSRC project grant (to PPD, DMG, EL and KSL). VA held postdoctoral fellowships from EMBO and HSFP. We thank M. Gatti, S. Bonaccorsi and M. Goldberg for antibodies, L. Capalbo for sharing data about Polo::GFP dynamics during mitosis, S. Hester for LC MS/MS and M. Savoian for his help with the spinning disc confocal and many helpful discussions. We also thank L.Capalbo, M. Savoian, T. Takeda and K. Verbrugghe for critical reading of the manuscript. Author Contributions: Conceived and designed the experiments: PD. Performed the experiments: MP PD VA WZ. Analyzed the data: DG PD VA. Contributed reagents/materials/analysis tools: DG EL PD KL. Wrote the paper: PD. The authors have declared that no competing interests exist.

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Supplemental Material - journal.pone.0000572.s006.AVI

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Supplemental Material - journal.pone.0000572.s009.AVI

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