Abstract
During mitosis and meiosis, the bipolar spindle facilitates chromosome segregation through microtubule sliding as well as microtubule growth and shrinkage1. Kinesin-14, one of the motors involved, causes spindle collapse in the absence of kinesin-5 (Refs 2, 3), participates in spindle assembly4 and modulates spindle length5. However, the molecular mechanisms underlying these activities are not known. Here, we report that Drosophila melanogaster kinesin-14 (Ncd) alone causes sliding of anti-parallel microtubules but locks together (that is, statically crosslinks) those that are parallel. Using single molecule imaging we show that Ncd diffuses along microtubules in a tail-dependent manner and switches its orientation between sliding microtubules. Our results show that kinesin-14 causes sliding and expansion of an anti-parallel microtubule array by dynamic interactions through the motor domain on the one side and the tail domain on the other. This mechanism accounts for the roles of kinesin-14 in spindle organization.
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References
Gadde, S. & Heald, R. Mechanisms and molecules of the mitotic spindle. Curr. Biol. 14, 797–805 (2004).
Sharp, D. J., Yu, K. R., Sisson, J. C., Sullivan, W. & Scholey, J. M. Antagonistic microtubule-sliding motors position mitotic centrosomes in Drosophila early embryos. Nature Cell Biol. 1, 51–54 (1999).
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).
Walczak, C. E., Verma, S. & Mitchison, T. J. XCTK2: a kinesin-related protein that promotes mitotic spindle assembly in Xenopus laevis egg extracts. J. Cell Biol. 136, 859–870 (1997).
Sharp, D. J. et al. Functional coordination of three mitotic motors in Drosophila embryos. Mol. Biol. Cell 11, 241–253 (2000).
Hildebrandt, E. R. & Hoyt, M. A. Mitotic motors in Saccharomyces cerevisiae. Biochim. Biophys. Acta 1496, 99–116 (2000).
Sharp, D. J., Rogers, G. C. & Scholey, J. M. Microtubule motors in mitosis. Nature 407, 41–47 (2000).
Chandra, R., Salmon, E. D., Erickson, H. P., Lockhart, A. & Endow, S. A. Structural and functional domains of the Drosophila ncd microtubule motor protein. J. Biol. Chem. 268, 9005–9013 (1993).
McDonald, H. B. & Goldstein, L. S. Identification and characterization of a gene encoding a kinesin-like protein in Drosophila. Cell 61, 991–1000 (1990).
McDonald, H. B., Stewart, R. J. & Goldstein, L. S. The kinesin-like ncd protein of Drosophila is a minus end-directed microtubule motor. Cell 63, 1159–1165 (1990).
Karabay, A. & Walker, R. A. Identification of microtubule binding sites in the Ncd tail domain. Biochemistry 38, 1838–1849 (1999).
Matthies, H. J., McDonald, H. B., Goldstein, L. S. & Theurkauf, W. E. Anastral meiotic spindle morphogenesis: role of the non-claret disjunctional kinesin-like protein. J. Cell Biol. 134, 455–464 (1996).
Kimble, M. & Church, K. Meiosis and early cleavage in Drosophila melanogaster eggs: effects of the claret-non-disjunctional mutation. J. Cell Sci. 62, 301–318 (1983).
Segbert, C. et al. KLP-18, a Klp2 kinesin, is required for assembly of acentrosomal meiotic spindles in Caenorhabditis elegans. Mol. Biol. Cell 14, 4458–4469 (2003).
Hatsumi, M. & Endow, S. A. Mutants of the microtubule motor protein, nonclaret disjunctional, affect spindle structure and chromosome movement in meiosis and mitosis. J. Cell Sci. 101, 547–559 (1992).
Saunders, W. S. & Hoyt, M. A. Kinesin-related proteins required for structural integrity of the mitotic spindle. Cell 70, 451–458 (1992).
Mountain, V. et al. The kinesin-related protein, HSET, opposes the activity of Eg5 and cross-links microtubules in the mammalian mitotic spindle. J. Cell Biol. 147, 351–366 (1999).
Goshima, G., Nedelec, F. & Vale, R. D. Mechanisms for focusing mitotic spindle poles by minus end-directed motor proteins. [see comment]. J. Cell Biol. 171, 229–240 (2005).
Endow, S. A., Chandra, R., Komma, D. J., Yamamoto, A. H. & Salmon, E. D. Mutants of the Drosophila ncd microtubule motor protein cause centrosomal and spindle pole defects in mitosis. J. Cell Sci. 107, 859–867 (1994).
Troxell, C. L. et al. pkl1+ and klp2+: Two kinesins of the Kar3 subfamily in fission yeast perform different functions in both mitosis and meiosis. Mol. Biol. Cell 12, 3476–3488 (2001).
Brust-Mascher, I. & Scholey, J. M. Microtubule flux and sliding in mitotic spindles of Drosophila embryos. Mol. Biol. Cell 13, 3967–3975 (2002).
Kapitein, L. C. et al. The bipolar mitotic kinesin Eg5 moves on both microtubules that it crosslinks. Nature 435, 114–118 (2005).
Hoyt, M. A., He, L., Totis, L. & Saunders, W. S. Loss of function of Saccharomyces cerevisiae kinesin-related CIN8 and KIP1 is suppressed by KAR3 motor domain mutations. Genetics 135, 35–44 (1993).
Oladipo, A., Cowan, A. & Rodionov, V. Microtubule motor Ncd induces sliding of microtubules in vivo. Mol. Biol. Cell 18, 3601–3606 (2007).
Foster, K. A. & Gilbert, S. P. Kinetic studies of dimeric Ncd: evidence that Ncd is not processive. Biochemistry 39, 1784–1791 (2000).
Case, R. B., Pierce, D. W., Hom-Booher, N., Hart, C. L. & Vale, R. D. The directional preference of kinesin motors is specified by an element outside of the motor catalytic domain. Cell 90, 959–966 (1997).
Furuta, K. & Toyoshima, Y. Y. Minus-end-directed motor Ncd exhibits processive movement that is enhanced by microtubule bundling in vitro. Curr. Biol. 18, 152–157 (2008).
Walczak, C. E., Vernos, I., Mitchison, T. J., Karsenti, E. & Heald, R. A model for the proposed roles of different microtubule-based motor proteins in establishing spindle bipolarity. Curr. Biol. 8, 903–913 (1998).
Kapitein, L. C. et al. Microtubule cross-linking triggers the directional motility of kinesin-5. J. Cell Biol. 182, 421–428 (2008).
van den Wildenberg, S. M. et al. The homotetrameric kinesin-5 KLP61F preferentially crosslinks microtubules into antiparallel orientations. Curr. Biol. 18, 1860–1864 (2008).
Cheerambathur, D. K., Brust-Mascher, I., Civelekoglu-Scholey, G. & Scholey, J. M. Dynamic partitioning of mitotic kinesin-5 cross-linkers between microtubule-bound and freely diffusing states. J. Cell Biol. 182, 429–436 (2008).
Uteng, M., Hentrich, C., Miura, K., Bieling, P. & Surrey, T. Poleward transport of Eg5 by dynein-dynactin in Xenopus laevis egg extract spindles. J. Cell Biol. 182, 715–726 (2008).
Helenius, J., Brouhard, G., Kalaidzidis, Y., Diez, S. & Howard, J. The depolymerizing kinesin MCAK uses lattice diffusion to rapidly target microtubule ends. Nature 441, 115–119 (2006).
Hyman, A. A. Preparation of marked microtubules for the assay of the polarity of microtubule-based motors by fluorescence. J. Cell Sci. Suppl. 14, 125–127 (1991).
Rogers, K. R. et al. KIF1D is a fast non-processive kinesin that demonstrates novel K-loop-dependent mechanochemistry. EMBO J. 20, 5101–5113 (2001).
Acknowledgements
We acknowledge the members of the Kasprzak and the Diez labs for comments on the manuscript as well as C. Bräuer and D. Naumburger for technical support. We thank C. Leduc, B. Nitzsche, C. Gell and J. Howard for fruitful discussion, F. Ruhnow for help with the microtubule tracking, as well as S. Bajer and E. Kocik for help with FPLC experiments. This work was supported by the German Federal Ministry of Education and Research (Grant 03 N 8712), the Polish Network for Mechanisms of Cell Motility, the Grant 2 P04C 131 29 and the Max-Planck-Society. G. Fink was supported by a fellowship from the Boehringer Ingelheim Foundation
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G.F., L.H., C.R., A.A.K. and S.D. designed the experiments; L.H. and C.R. performed initial experiments; G.F. performed the presented experiments and analysed the data; L.H. and K.J.S. generated the Ncd proteins; G.F and S.D. wrote the manuscript; A.A.K. and S.D. initiated the research and supervised the work. All authors discussed the results and commented on the manuscript.
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Fink, G., Hajdo, L., Skowronek, K. et al. The mitotic kinesin-14 Ncd drives directional microtubule–microtubule sliding. Nat Cell Biol 11, 717–723 (2009). https://doi.org/10.1038/ncb1877
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DOI: https://doi.org/10.1038/ncb1877
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