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The cilia protein IFT88 is required for spindle orientation in mitosis

Nature Cell Biology volume 13, pages 461–468 (2011)Cite this article

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Abstract

Cilia dysfunction has long been associated with cyst formation and ciliopathies1. More recently, misoriented cell division has been observed in cystic kidneys2, but the molecular mechanism leading to this abnormality remains unclear. Proteins of the intraflagellar transport (IFT) machinery are linked to cystogenesis and are required for cilia formation in non-cycling cells3,4. Several IFT proteins also localize to spindle poles in mitosis5,6,7,8, indicating uncharacterized functions for these proteins in dividing cells. Here, we show that IFT88 depletion induces mitotic defects in human cultured cells, in kidney cells from the IFT88 mouse mutant Tg737orpk and in zebrafish embryos. In mitosis, IFT88 is part of a dynein1-driven complex that transports peripheral microtubule clusters containing microtubule-nucleating proteins to spindle poles to ensure proper formation of astral microtubule arrays and thus proper spindle orientation. This work identifies a mitotic mechanism for a cilia protein in the orientation of cell division and has important implications for the etiology of ciliopathies.

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Figure 1: IFT88 depletion leads to mitotic defects in HeLa cells, kidney cells from the Tg737orpk mouse mutant and zebrafish.
Figure 2: IFT88 depletion disrupts astral microtubules and the spindle pole localization of proteins involved in microtubule nucleation in HeLa cells.
Figure 3: IFT88 is required for the movement of peripheral microtubule clusters containing microtubule-nucleating components towards spindle poles in LLC-PK1 cells stably expressing GFP– α -tubulin.
Figure 4: IFT88 moves towards spindle poles and requires microtubules for its spindle pole localization.
Figure 5: IFT88 is part of a dynein1-driven transport complex in mitosis.

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Acknowledgements

We thank G. Pazour, G. Witman and P. Wadsworth for thoughtful discussions on this work, and S. Redick for assistance with microscopy. We are particularly grateful to L. Covassin-Barberis in N.L. laboratory and N.L. Adkins in C. Peterson’s laboratory for guidance on zebrafish experimental work and gel-filtration experiments, respectively. We thank G. Pazour for the gift of IFT88 antibody and GFP–IFT88 LLC-PK1, Flag–IFT52 IMCD stable cell lines, P. Wadsworth for the GFP– α -tubulin LLC-PK1 cell line and C. Desdouets, P. Denoulet and C. Janke for their gift of antibodies specific to IFT88 and polyglutamylated tubulin, respectively. Core resources supported by the Diabetes Endocrinology Research Center grant DK32520 were used; S.D. is a member of the UMass DERC (DK32520). This work was supported by financial support from the National Institutes of Health (GM51994) to S.D. and the Polycystic Kidney Disease Foundation to B.D.

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  1. Program in Molecular Medicine, University of Massachusetts Medical School, 373 Plantation Street, Suite 210, Worcester, Massachusetts 01605, USA

    Benedicte Delaval, Alison Bright & Stephen Doxsey

  2. Program in Gene Function and Expression, University of Massachusetts Medical School, Worcester, Massachusetts 01605, USA

    Nathan D. Lawson

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  1. Benedicte Delaval
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B.D. and S.D. wrote the manuscript. B.D. conceived and planned the experimental work. B.D. and A.B. carried out the experimental work and analysed the data. N.D.L. provided the zebrafish facility and helped plan and guide the zebrafish experimental work.

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Correspondence to Stephen Doxsey.

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Delaval, B., Bright, A., Lawson, N. et al. The cilia protein IFT88 is required for spindle orientation in mitosis. Nat Cell Biol 13, 461–468 (2011). https://doi.org/10.1038/ncb2202

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