Abstract
The XMAP215/ch-TOG/Msps family of microtubule-associated proteins (MAPs) promote microtubule growth in vitro and are concentrated at centrosomes in vivo. We show here that Msps (mini-spindles protein) interacts with the centrosomal protein D-TACC, and that this interaction strongly influences microtubule behaviour in Drosophila embryos. If D-TACC levels are reduced, Msps does not concentrate at the centrosomes efficiently and the centrosomal microtubules appear to be destabilized. If D-TACC levels are increased, both D-TACC and Msps accumulate around the centrosomes/spindle poles, and the centrosomal microtubules appear to be stabilized. We show that the interaction between D-TACC and Msps is evolutionarily conserved. We propose that D-TACC and Msps normally cooperate to stabilize centrosomal microtubules by binding to their minus ends and binding to their plus ends as they grow out from the centrosome.
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Acknowledgements
We thank H. Ohkura and F. Cullen for sharing results before publication and for supplying anti-Msps anti-serum and the mspsMJ15 allele. We thank B. Williams and M. Goldberg who provided us with their unpublished d-taccstella alleles. We also thank T. Hyman and D. Compton for providing us with anti-ch-TOG antibodies, and J. Kilmartin for advice about sample preparation for mass spectroscopy. We thank members of the laboratory for comments on the manuscript. This work was supported by a Wellcome Trust Senior Fellowship in Basic Biomedical Sciences (to J.W.R.), an MRC studentship (to M.J.L.), a Wellcome Trust Prize Studentship (to F.G.) and the Medical Research Council (S.Y.P.-C.).
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Movie 1
The behaviour of a D-TACC–GFP fusion protein in a living embryo. Note how small dots of the fusion protein can be observed oscillating to and fro from the centrosomal region, as though binding to the plus ends of the centrosomal microtubules. (MOV 7992 kb)
Movie 2
The behaviour of a Msps–GFP fusion protein in a living embryo. Note how small dots of the fusion protein can be observed oscillating to and fro from the centrosomal region, as though binding to the plus ends of the centrosomal microtubules. (MOV 3252 kb)
Figure S1
The behaviour of D–TACC (top left panel), Msps (bottom left panel), microtubules (middle panel) and DNA (right panel) in embryos treated with taxol before fixation. (PDF 288 kb)
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Lee, M., Gergely, F., Jeffers, K. et al. Msps/XMAP215 interacts with the centrosomal protein D-TACC to regulate microtubule behaviour. Nat Cell Biol 3, 643–649 (2001). https://doi.org/10.1038/35083033
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DOI: https://doi.org/10.1038/35083033
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