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A cytoplasmic dynein tail mutation impairs motor processivity

Nature Cell Biology volume 12, pages 1228–1234 (2010)Cite this article

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Abstract

Mutations in the tail of the cytoplasmic dynein molecule have been reported to cause neurodegenerative disease in mice. The mutant mouse strain Legs at odd angles (Loa) has impaired retrograde axonal transport, but the molecular deficiencies in the mutant dynein molecule, and how they contribute to neurodegeneration, are unknown. To address these questions, we purified dynein from wild-type mice and the Legs at odd angles mutant mice. Using biochemical, single-molecule, and live-cell-imaging techniques, we find a marked inhibition of motor run-length in vitro and in vivo, and significantly altered motor domain coordination in the dynein from mutant mice. These results suggest a potential role for the dynein tail in motor function, and provide direct evidence for a link between single-motor processivity and disease.

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Figure 1: Purification and biochemical analysis of wild-type and mutant cytoplasmic dynein.
Figure 2: Single-molecule dynamics of cytoplasmic dynein isolated from wild-type and mutant mice.
Figure 3: Optical-trap analysis of wild-type and mutant cytoplasmic dynein dynamics in single- and multi-motor regions.
Figure 4: Analysis of retrograde transport of lysosomes in wild-type and mutant axons, with theoretical comparison.
Figure 5: Biophysical and biochemical evidence of altered motor coordination in mutant dynein.

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Acknowledgements

We thank R. McKenney for help with the quantum dot assays and useful discussion, and M. Vershinnin for developing the tracking program. This work was supported by NIH grants GM47434 (to R.B.V.), RO1GM070676 (to S.P.G.) and GM008798-09 (to K.M.O.M.), AHA grant 825278F (to J.X.), and the Columbia University Motor Neuron Center.

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Author notes

  1. Kassandra M. Ori-McKenney and Jing Xu: These authors contributed equally to this work.

  2. Steven P. Gross and Richard B. Vallee: Co-senior authors.

Authors and Affiliations

  1. Department of Pathology and Cell Biology, Columbia University., New York, 10032, NY, USA

    Kassandra M. Ori-McKenney & Richard B. Vallee

  2. Department of Biological Sciences, Columbia University, New York, 10027, NY, USA

    Kassandra M. Ori-McKenney & Richard B. Vallee

  3. Department of Developmental and Cell Biology, University of California, Irvine, 92697, CA, USA

    Jing Xu & Steven P. Gross

Authors
  1. Kassandra M. Ori-McKenney
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  2. Jing Xu
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Contributions

K.M.O.M., J.X., S.P.G. and R.B.V. designed the research. K.M.O.M. and J.X. performed experiments and analysed data. K.M.O.M., J.X., S.P.G. and R.B.V. wrote the paper.

Corresponding authors

Correspondence to Steven P. Gross or Richard B. Vallee.

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The authors declare no competing financial interests.

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Ori-McKenney, K., Xu, J., Gross, S. et al. A cytoplasmic dynein tail mutation impairs motor processivity. Nat Cell Biol 12, 1228–1234 (2010). https://doi.org/10.1038/ncb2127

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