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Chromosome- and spindle-pole-derived signals generate an intrinsic code for spindle position and orientation

Nature Cell Biology volume 14, pages 311–317 (2012)Cite this article

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Abstract

Mitotic spindle positioning by cortical pulling forces1 defines the cell division axis and location2, which is critical for proper cell division and development3. Although recent work has identified developmental and extrinsic cues that regulate spindle orientation4,5,6, the contribution of intrinsic signals to spindle positioning and orientation remains unclear. Here, we demonstrate that cortical force generation in human cells is controlled by distinct spindle-pole- and chromosome-derived signals that regulate cytoplasmic dynein localization. First, dynein exhibits a dynamic asymmetric cortical localization that is negatively regulated by spindle-pole proximity, resulting in spindle oscillations to centre the spindle within the cell. We find that this signal comprises the spindle-pole-localized polo-like kinase (Plk1), which regulates dynein localization by controlling the interaction between dynein–dynactin and its upstream cortical targeting factors NuMA and LGN. Second, a chromosome-derived RanGTP gradient restricts the localization of NuMA–LGN to the lateral cell cortex to define and maintain the spindle orientation axis. RanGTP acts in part through the nuclear localization sequence of NuMA to locally alter the ability of NuMA–LGN to associate with the cell cortex in the vicinity of chromosomes. We propose that these chromosome- and spindle-pole-derived gradients generate an intrinsic code to control spindle position and orientation.

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Figure 1: Dynein and dynactin localize asymmetrically to the cell cortex during metaphase.
Figure 2: Plk1 negatively regulates the localization of cortical dynein.
Figure 3: The chromosome-derived Ran gradient negatively regulates LGN localization.
Figure 4: The Ran gradient regulates the association of the LGN–NuMA complex with membranes.

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Acknowledgements

We thank P. Meraldi, D. Pellman, D. Yarar, K. Ribbeck and members of the Cheeseman and Hochwagen laboratories for discussions and critical reading of the manuscript, and T. DiCesare for help with generating the movie. We also thank D. Compton, T. Hyman-Mitocheck, C. Basilico and A. C. Groen for reagents. This work was supported by awards to I.M.C. from the Massachusetts Life Sciences Center, the Searle Scholars Program and the Human Frontiers Science Foundation, a grant from the National Institute of General Medical Sciences (GM088313) and a Research Scholar Grant (121776) from the American Cancer Society. I.M.C. is a Thomas D. and Virginia W. Cabot Career Development Professor of Biology. T.K. is supported by a long-term fellowship of the Human Frontiers Science Program.

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  1. Whitehead Institute, Nine Cambridge Center, Cambridge, Massachusetts 02142, USA

    Tomomi Kiyomitsu & Iain M. Cheeseman

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  1. Tomomi Kiyomitsu
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T.K. and I.M.C. designed the experiments. T.K. carried out all of the experiments. I.M.C. assisted with some experiments including protein purification and mass spectrometry. T.K. and I.M.C. analysed the data and wrote the manuscript.

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Correspondence to Iain M. Cheeseman.

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Kiyomitsu, T., Cheeseman, I. Chromosome- and spindle-pole-derived signals generate an intrinsic code for spindle position and orientation. Nat Cell Biol 14, 311–317 (2012). https://doi.org/10.1038/ncb2440

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