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A more recent version of this article appeared on April 1, 2004
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Submitted on November 24, 2003
Revised on January 7, 2004
Accepted on January 7, 2004
1 Laboratory of Cell Regulation, Cancer Research UK, London Research Institute, Lincoln’s Inn Fields Laboratories, 44 Lincoln’s Inn Fields, London WC2A 3PX, UK
2 Laboratory of Cell Regulation, Cancer Research UK, London Research Institute, Lincoln’s Inn Fields Laboratories, 44 Lincoln’s Inn Fields, London WC2A 3PX, UK, Room539, Whitehead Institute, nine Cambridge Center, Cambridge, MA 02142, U. S. A.
3 Laboratory of Cell Regulation, Cancer Research UK, London Research Institute, Lincoln’s Inn Fields Laboratories, 44 Lincoln’s Inn Fields, London WC2A 3PX, UK, Centro de Biología Molecular "Severo Ochoa", Universidad Autónoma de Madrid, 28049 Cantoblanco, Madrid, Spain
* Corresponding author. E-mail address: toda{at}cancer.org.uk.
The Dis1/TOG family plays a pivotal role in microtubule organization. In fission yeast Alp14 and Dis1 share an essential function in bipolar spindle formation. Here we characterize Alp7, a novel coiled-coil protein that is required for organization of bipolar spindles. Both Alp7 and Alp14 colocalize to the spindle pole body (SPB) and mitotic spindles. Alp14 localization to these sites is fully dependent upon Alp7. Conversely in the absence of Alp14, Alp7 localizes to the SPBs, but not mitotic spindles. Alp7 forms a complex with Alp14, where the C-terminal region of Alp14 interacts with the coiled-coil domain of Alp7. Intriguingly this Alp14 C-terminus is necessary and sufficient for mitotic spindle localization. Overproduction of either full-length or coiled-coil region of Alp7 results in abnormal V-shaped spindles and stabilization of interphase microtubules, which is induced independent of Alp14. Alp7 may be a functional homologue of animal TACC. Our results shed light on an interdependent relationship between Alp14/TOG and Alp7. We propose a two-step model that accounts for the recruitment of Alp7 and Alp14 to the SPB and microtubules.
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