The positioning of centrosomes, or microtubule-organizing centres, within cells plays a critical part in animal development. Here we show that, in Drosophila embryos undergoing mitosis, the positioning of centrosomes within bipolar spindles and between daughter nuclei is determined by a balance of opposing forces generated by a bipolar kinesin motor, KLP61F, that is directed to microtubule plus ends, and a carboxy-terminal kinesin motor, Ncd, that is directed towards microtubule minus ends. This activity maintains the spacing between separated centrosomes during prometaphase and metaphase, and repositions centrosomes and daughter nuclei during late anaphase and telophase. Surprisingly, we do not observe a function for KLP61F in the initial separation of centrosomes during prophase. Our data indicate that KLP61F and Ncd may function by crosslinking and sliding antiparallel spindle microtubules in relation to one another, allowing KLP61F to push centrosomes apart and Ncd to pull them together.
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Rappaport, R. in Cytokinesis in Animal Cells (eds Barlow, P. W., Bard, J. B. L., Green, P. B. & Kirk, D. L.) 1–386 (Cambridge Univ. Press, Cambridge, 1996).
Sullivan, W. & Theurkauf, W. E. The cytoskeleton and morphogenesis of the early Drosophila embryo. Curr. Opin. Cell Biol. 7, 18–22 (1995).
Hoyt, M. A. & Geiser, J. R. Genetic analysis of the mitotic spindle. Annu. Rev. Genet. 30, 7– 33 (1996).
Vale, R. D. & Fletterick, R. J. The design plan of kinesin motors. Annu. Rev. Cell Dev. Biol. 13, 745 –777 (1997).
Enos, A. P. & Morris, N. R. Mutation of a gene that encodes a kinesin-like protein blocks nuclear division in Aspergillus nidulans. Cell 60, 1019– 1027 (1990).
Roof, D. M., Meluh, P. B. & Rose, M. D. Kinesin-related proteins required for assembly of the mitotic spindle. J. Cell. Biol. 118, 95–108 (1992).
Hoyt, M. A., He, L., Loo K. K. & Saunders, W. S. Two Saccharomyces cerevisiae kinesin-related gene-products required for mitotic spindle assembly. J. Cell. Biol. 118, 109–120 ( 1992).
Saunders, W. S. & Hoyt, M. A. Kinesin-related proteins required for structural integrity of the mitotic spindle. Cell 70, 451–458 ( 1992).
Hagan, I. & Yanagida, M. Novel potential mitotic motor protein encoded by the fission yeast Cut7+ gene. Nature 347 , 563–566 (1990).
Hagan, I. & Yanagida, M. Kinesin-related Cut7 protein associates with mitotic and meiotic spindles in fission yeast. Nature 356, 74–76 (1992).
Straight, A. F., Sedat, J. W. & Murray, A. W. Time-lapse microscopy reveals unique roles for kinesins during anaphase in budding yeast. J. Cell. Biol. 143 , 687–694 (1998).
Sawin, K. E., Leguellec, K., Phillipe, M. & Mitchison, T. J. Mitotic spindle organization by a plus-end-directed microtubule motor. Nature 359, 540–543 ( 1992).
Heck, M. M. et al. The kinesin-like protein KLP61F is essential for mitosis in Drosophila. J. Cell Biol. 123, 665– 679 (1993).
Cole, D. G., Saxton, W. M., Sheehan, K. B. & Scholey, J. M. A ‘slow’, homotetrameric, plus-end directed kinesin purified from Drosophila embryos. J. Biol. Chem. 269, 22913–22916 (1994).
Kashina, A.S. et al. A bipolar kinesin. Nature 379, 270–272 (1996).
Kashina, A. S., Scholey, J. M., Leszyk, J. D. & Saxton, W. M. An essential bipolar mitotic motor. Nature 384, 225 (1996).
Sharp, D.J. et al. The bipolar kinesin, KLP61F, crosslinks microtubules within interpolar microtubule bundles of Drosophila embryonic mitotic spindles . J. Cell. Biol. 144, 125– 138 (1999).
Blangy, A. et al. Phosphorylation by p34(Cdc2) regulates spindle association of human Eg5, a kinesin-related motor essential for bipolar spindle formation in vivo. Cell 83, 1159– 1169 (1995).
McDonald, H. B., Stewart, R. J. & Goldstein, L. S. B. The kinesin-like Ncd protein of Drosophila is a minus-end directed microtubule motor. Cell 63, 1159–1165 (1990).
Walker, R. A., Salmon, E. D. & Endow, S. A. The Drosophila claret segregation protein is a minus-end directed motor molecule. Nature 347, 780–782 (1990).
Matthies, H. J., McDonald, H. B., Goldstein, L. S. B. & Theurkauf, W. E. Anastral meiotic spindle morphogenesis: role of the Non-claret Disjunctional kinesin-like protein. J.Cell Biol. 134, 455–464 (1996).
Endow, S. A. & Komma, D. J. Centrosome and spindle function of the Drosophila Ncd microtubule motor visualized in live embryos using Ncd-GFP fusion proteins. J. Cell Sci. 109, 2429–2442 (1996).
O’Connell, M. J., Meluh, P. B., Rose, M. D. & Morris, M. R. Suppression of the bimC mitotic spindle defect by deletion of klpA, a kar3 related kinesin like protein in Aspergillus nidulans. J. Cell Biol. 120, 153–162 (1993).
Pidoux, A. L., LeDizet, M. & Cande, W. Z. Fission yeast pkl1 is a kinesin-related protein involved in mitotic spindle function. Mol. Biol. Cell. 7, 1639–1655 (1996).
Saunders, W., Lengyel, V. & Hoyt, M. A. Mitotic spindle function in Saccharomyces cerevisiae requires a balance between different types of kinesin-related motors . Mol. Biol. Cell. 8, 1025– 1033 (1997).
Su, T. T. et al. Exit from mitosis in Drosophila syncytial embryos requires proteolysis and cyclin degradation and is associated with localized dephosphorylation . Genes Dev. 12, 1495–1503 (1998).
Lewis, E. B. & Gancarella, W. Claret and non-disjunction. Genetics 37, 600–601 ( 1952).
Vaisberg, E. A., Koonce, M. P. & McIntosh, J. R. Cytoplasmic dynein plays a role in mammalian mitotic spindle formation. J. Cell Biol. 123, 849 –858 (1993).
Saunders, W. S., Koshland, D., Eshel, D., Gibbons, I. R. & Hoyt, M. A. Saccharomyces cerevisiae kinesin- and dynein-related proteins required for anaphase chromosome segregation. J. Cell Biol. 128, 617–624 ( 1995).
McIntosh, J. R., Hepler, P. K. & Van Wie, D. G. Model for mitosis. Nature 224, 659–663 (1969).
Francis-Lang, H., Minden, J., Sullivan, W. & Oegema, K. Live confocal analysis with fluorescently labeled proteins. Methods Mol. Biol. 122, 223–237 (1995).
Kellogg, D. R. et al. Studies on the centrosome and cytoplasmic organization in the early Drosophila embryo. Cold Spring Harb. Symp. Quant. Biol. 56, 649–662 ( 1991).
This work was supported by grants from the NIH (to J.M.S. and W.S.) and NIH postdoctoral fellowships (to D.J.S. and J.C.S.). We thank S. Hawley and members of his laboratory for help with this study and for providing the Cand fly stocks; R. Saint for providing the histone–GFP line; G. Rogers for Fig. 4 and his intellectual contribution to this work; and the other members of the Scholey and Sullivan laboratories for assistance. The anti-KLP61F antibody used here was made in collaboration with T. Mitchison and we thank him for his continuing interest and for discussions about mitosis.
Correspondence and requests for materials should be addressed to J.M.S.
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Sharp, D., Yu, K., Sisson, J. et al. Antagonistic microtubule-sliding motors position mitotic centrosomes in Drosophila early embryos. Nat Cell Biol 1, 51–54 (1999). https://doi.org/10.1038/9025
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