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Structural basis for midbody targeting of spastin by the ESCRT-III protein CHMP1B

Nature Structural & Molecular Biology volume 15, pages 12781286 (2008) | Download Citation

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Abstract

The endosomal sorting complex required for transport (ESCRT) machinery, including ESCRT-III, localizes to the midbody and participates in the membrane-abscission step of cytokinesis. The ESCRT-III protein charged multivesicular body protein 1B (CHMP1B) is required for recruitment of the MIT domain–containing protein spastin, a microtubule-severing enzyme, to the midbody. The 2.5-Å structure of the C-terminal tail of CHMP1B with the MIT domain of spastin reveals a specific, high-affinity complex involving a noncanonical binding site between the first and third helices of the MIT domain. The structural interface is twice as large as that of the MIT domain of the VPS4–CHMP complex, consistent with the high affinity of the interaction. A series of unique hydrogen-bonding interactions and close packing of small side chains discriminate against the other ten human ESCRT-III subunits. Point mutants in the CHMP1B binding site of spastin block recruitment of spastin to the midbody and impair cytokinesis.

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Acknowledgements

We thank N. Elia for discussions, the staff of SER-CAT for user support at the Advance Photon Source (APS), C.-R. Chang for technical assistance, E. Tyler for generating Figure 6 and D. Davies for comments on the manuscript. Use of the APS was supported by the US Department of Energy, Basic Energy Sciences, Office of Science, under Contract No.W-31-109-Eng-38. This project was funded by the Intramural Research Programs of US National Institute of Diabetes and Digestive and Kidney Diseases, US National Institute of Neurological Disorders and Stroke and the US National Institute of Child Health and Human Development, and the Bench-to-Bedside program of the US National Institutes of Health (NIH).

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Affiliations

  1. Laboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, US Department of Health and Human Services, Bethesda, Maryland 20892, USA.

    • Dong Yang
    •  & James H Hurley
  2. Cellular Neurology Unit, Neurogenetics Branch, National Institute of Neurological Disorders and Stroke, National Institutes of Health, US Department of Health and Human Services, Bethesda, Maryland 20892, USA.

    • Neggy Rismanchi
    • , Benoît Renvoisé
    •  & Craig Blackstone
  3. Cell Biology and Metabolism Program, National Institute of Child Health and Human Development, National Institutes of Health, US Department of Health and Human Services, Bethesda, Maryland 20892, USA.

    • Jennifer Lippincott-Schwartz

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Contributions

D.Y. carried out binding experiments, crystallization and structure determination; N.R. and B.R. carried out knockdown and cell-imaging experiments; J.L.-S. interpreted data; C.B. carried out yeast two-hybrid experiments; C.B. and J.H.H. designed research, interpreted data and wrote the paper.

Corresponding authors

Correspondence to Craig Blackstone or James H Hurley.

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https://doi.org/10.1038/nsmb.1512

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