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Structural basis for ESCRT-III protein autoinhibition

Nature Structural & Molecular Biology volume 16, pages 754–762 (2009)Cite this article

Abstract

Endosomal sorting complexes required for transport-III (ESCRT-III) subunits cycle between two states: soluble monomers and higher-order assemblies that bind and remodel membranes during endosomal vesicle formation, midbody abscission and enveloped virus budding. Here we show that the N-terminal core domains of increased sodium tolerance-1 (IST1) and charged multivesicular body protein-3 (CHMP3) form equivalent four-helix bundles, revealing that IST1 is a previously unrecognized ESCRT-III family member. IST1 and its ESCRT-III binding partner, CHMP1B, both form higher-order helical structures in vitro, and IST1-CHMP1 interactions are required for abscission. The IST1 and CHMP3 structures also reveal that equivalent downstream α5 helices can fold back against the core domains. Mutations within the CHMP3 core–α5 interface stimulate the protein's in vitro assembly and HIV-inhibition activities, indicating that dissociation of the autoinhibitory α5 helix from the core activates ESCRT-III proteins for assembly at membranes.

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Figure 1: CHMP3, CHMP38–222 and IST1NTD are monomers in solution.
Figure 2: Structures of IST1NTD and CHMP3.
Figure 3: CHMP3 crystal packing interactions.
Figure 4: Mutational analyses of IST1–CHMP1B interactions.
Figure 5: Requirement for IST1–CHMP1 interactions during abscission.
Figure 6: IST1NTD and CHMP1B tube assembly.
Figure 7: CHMP3 activation in vitro.
Figure 8: CHMP3 activation in vivo.

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Acknowledgements

Portions of this research were carried out at the Stanford Synchrotron Radiation Light Source (SSRL), a national user facility operated by Stanford University on behalf of the US Department of Energy, Office of Basic Energy Sciences. The SSRL Structural Molecular Biology Program is supported by the Department of Energy, Office of Biological and Environmental Research and by the US National Institutes of Health (NIH), National Center for Research Resources, Biomedical Technology Program, and the National Institute of General Medical Sciences. We thank M. Babst for helpful discussions. This work was supported by NIH grants AI051174 (W.I.S.) and GM082545 (W.I.S. and C.P.H.).

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  1. Monika Bajorek, Heidi L Schubert and John McCullough: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Biochemistry, University of Utah, Salt Lake City, Utah, USA

    Monika Bajorek, Heidi L Schubert, John McCullough, Charles Langelier, Debra M Eckert, William-May B Stubblefield, Nathan T Uter, David G Myszka, Christopher P Hill & Wesley I Sundquist

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Correspondence to Christopher P Hill or Wesley I Sundquist.

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Bajorek, M., Schubert, H., McCullough, J. et al. Structural basis for ESCRT-III protein autoinhibition. Nat Struct Mol Biol 16, 754–762 (2009). https://doi.org/10.1038/nsmb.1621

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