Antagonistic microtubule-sliding motors position mitotic centrosomes in Drosophila early embryos

Abstract

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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Figure 1: Centrosomes within bipolar spindles separate but then collapse together following the inhibition of the bipolar kinesin KLP61F.
Figure 2: Mitotic centrosomes associated with adjacent spindles interact through microtubule bundles to slide together and form large arrays of collapsed mono-asters in the absence of KLP61F function.
Figure 3: Positioning of mitotic centrosomes and daughter nuclei involves an interaction between bipolar kinesins and C-terminal kinesins.
Figure 4: The relative positioning of mitotic centrosomes is determined by a balance of microtubule–microtubule sliding motors.

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Acknowledgements

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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Correspondence to Jonathan M. Scholey.

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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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