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A role for the lissencephaly gene LIS1 in mitosis and cytoplasmic dynein function

Nature Cell Biology volume 2, pages 784–791 (2000)Cite this article

Abstract

Mutations in the LIS1 gene cause gross histological disorganization of the developing human brain, resulting in a brain surface that is almost smooth. Here we show that LIS1 protein co-immunoprecipitates with cytoplasmic dynein and dynactin, and localizes to the cell cortex and to mitotic kinetochores, which are known sites for binding of cytoplasmic dynein. Overexpression of LIS1 in cultured mammalian cells interferes with mitotic progression and leads to spindle misorientation. Injection of anti-LIS1 antibody interferes with attachment of chromosomes to the metaphase plate, and leads to chromosome loss. We conclude that LIS1 participates in a subset of dynein functions, and may regulate the division of neuronal progenitor cells in the developing brain.

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Figure 1: Co-immunoprecipitation of cytoplasmic dynein and dynactin with LIS1.
Figure 2: Overexpression of LIS1 perturbs mitotic progression.
Figure 3: Phenotypic effects of LIS1 overexpression.
Figure 4: Effects of LIS1 overexpression on dynactin distribution.
Figure 5: Effects of anti-LIS1 antibody on the behaviour of mitotic chromosomes.
Figure 6: Subcellular localization of LIS1.
Figure 7: Overexpression of LIS1 displaces p150Glued from microtubule plus ends.
Figure 8: Potential functions of LIS1 in mitosis and brain development.

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Acknowledgements

We thank S. Lambert and E. Hinchcliffe for critical reading of the manuscript, and G. Sluder for helpful comments. We also thank O. Reiner, K. Pfister, H. Goodson, A. Mikami, D. Meyer and P. Denoulet for antibody reagents, D. Shima and G. Warren for GFP-N-acetyl glucosamine transferase cDNA, and J. De Mey for the MDCK cell line. This work was supported by NIH grants GM47434 and HD61982, the March of Dimes Birth Defects Foundation, the Human Frontier Science Program, and the H. Arthur Smith Charitable Foundation.

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  1. Kevin T. Vaughan

    Present address: Department of Biological Sciences, University of Notre Dame, Notre Dame, Indiana, 46556, USA

Authors and Affiliations

  1. Department of Cell Biology University of Massachusetts Medical School, 377 Plantation Street, Worcester, 01605, Massachusetts, USA

    Nicole E. Faulkner, Denis L. Dujardin, Chin-Yin Tai & Richard B. Vallee

  2. Physiology, University of Massachusetts Medical School, 377 Plantation Street, Worcester, 01605, Massachusetts, USA

    Christopher B. O'Connell & Yu-li Wang

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Supplementary information

Movie 1

Mitosis in control cells. Two control NRK cells, each injected with a pre-immune immunoglobulin G (IgG) fraction, were monitored as they progressed through mitosis. The sequence begins during early prometaphase.Mitosis proceeded rapidly and almost synchronously in the two cells. Times from nuclear-envelope breakdown (NEB) to anaphase onset are 21 min (left cell) and 15 min (right cell). Total elapsed time is 29 min. (MOV 172 kb)

Movie 2

Mitosis in an anti-LIS1-injected cell. The central cell was injected with anti-LIS1 antibody. The sequence begins at NEB. After formation of the metaphase plate, a prolonged delay is observed while several chromosomes remain unaligned. Time from NEB to anaphase onset is 48 min. Total elapsed time is 57 min. (MOV 768 kb)

Movie 3

Mitosis in an anti-LIS1-injected cell. The central cell was injected with anti-LIS1 antibody. The sequence begins at NEB. After a prolonged delay while individual chromosomes remain unaligned at metaphase plate, the cell proceeds through anaphase. The unattached chromosome remains behind and forms a micronucleus. Time from NEB to anaphase onset is 77 min. Total elapsed time is 100 min. (MOV 1497 kb)

Movie 4

Mitosis in an anti-LIS1-injected cell. Similar to Movie 3, except that the sequence begins during early prometaphase. Time from NEB to anaphase onset is 59 min. Total elapsed time is 73 min. A control mitotic cell can be seen on the left. (MOV 1721 kb)

Movie 5

Mitosis in a cell injected with antibody against cytoplasmic dynein. The upper-right cell was injected with an antibody against the cytoplasmic- dynein intermediate chain. The sequence begins at NEB. Note the repeated alignment and detechment of individual chromosomes during the prolonged delay. Time from NEB to anaphase onset is 90 min. Total elapsed time is 153 min. (MOV 1126 kb)

Five time-lapse sequences are shown of antibody-injected normal rat kidney (NRK) cells. Chromosomes were visualized by prestaining with Hoechst 33258, and were more easily observed in a darkened room.

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Faulkner, N., Dujardin, D., Tai, CY. et al. A role for the lissencephaly gene LIS1 in mitosis and cytoplasmic dynein function. Nat Cell Biol 2, 784–791 (2000). https://doi.org/10.1038/35041020

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