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Structure of the ESCRT-II endosomal trafficking complex

Nature volume 431, pages 221–225 (2004)Cite this article

Abstract

The multivesicular-body (MVB) pathway delivers transmembrane proteins and lipids to the lumen of the endosome. The multivesicular-body sorting pathway has crucial roles in growth-factor-receptor downregulation1, developmental signalling2,3,4, regulation of the immune response5 and the budding of certain enveloped viruses such as human immunodeficiency virus6. Ubiquitination is a signal for sorting into the MVB pathway7,8, which also requires the functions of three protein complexes, termed ESCRT-I, -II and -III (endosomal sorting complex required for transport)7,9,10. Here we report the crystal structure of the core of the yeast ESCRT-II complex, which contains one molecule of the Vps protein Vps22, the carboxy-terminal domain of Vps36 and two molecules of Vps25, and has the shape of a capital letter ‘Y’. The amino-terminal coiled coil of Vps22 and the flexible linker leading to the ubiquitin-binding NZF domain of Vps36 both protrude from the tip of one branch of the ‘Y’. Vps22 and Vps36 form nearly equivalent interactions with the two Vps25 molecules at the centre of the ‘Y’. The structure suggests how ubiquitinated cargo could be passed between ESCRT components of the MVB pathway through the sequential transfer of ubiquitinated cargo from one complex to the next.

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Figure 1: Structure of the ESCRT-II complex.
Figure 2: Winged helix folds in ESCRT-II.
Figure 3: ESCRT-II complex assembly and subunit interactions.
Figure 4: Mutational analysis of ESCRT-II subunit interfaces.

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Acknowledgements

We thank P. P. di Fiore, M. Babst and B. Wendland for helpful discussions; M. Tabuchi for the Flag tag construct; and we acknowledge the use of the SER-CAT beamline at the APS, ANL and beamline 9-2 at the Stanford Synchrotron Radiation Laboratory. Use of the Advanced Photon Source was supported by the US Department of Energy, Basic Energy Sciences, Office of Science. The Stanford Synchrotron Radiation Laboratory, a national user facility operated by Stanford University on behalf of the US Department of Energy, is supported by the Department of Energy, Office of Biological and Environmental Research, and by the National Institutes of Health, National Center for Research Resources, Biomedical Technology Program and the National Institute of General Medical Sciences. S.D.E. is supported as an Investigator of the Howard Hughes Medical Institute.

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Authors and 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, 20892-0580, USA

    Aitor Hierro, Jaewon Kim, Gali Prag & James H. Hurley

  2. Department of Cellular and Molecular Medicine and Department of Chemistry and Biochemistry and Howard Hughes Medical Institute, University of California at San Diego, 9500 Gilman Drive, La Jolla, California, 92093-0668, USA

    Ji Sun, Alexander S. Rusnak & Scott D. Emr

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Correspondence to James H. Hurley.

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Supplementary information

Supplementary Figure 1

Structures of ESCRT-II subunits. a. Structure of Vps22. b. Structure of Vps25. c. Structure of Vps36. (JPG 38 kb)

Supplementary Figure 2

Sequence conservation in ESCRT-II. a, Structure-based alignment of Vps22 and homologues with amino acids highlighted in red (identical in all species), and orange (conserved in at least four of five species shown). b, Vps25 and homologues; c, Vps36 C-terminal domain and homologues. (JPG 384 kb)

Supplementary Figure 3

Trafficking of ubiquitinated membrane proteins by ESCRTs. ESCRT-I binds Ub-cargo through the UEV domain of Vps23 23. The second NZF domain of Vps36 binds Ub-cargo (JPG 113 kb)

Supplementary Table 1

Crystallographic data Collection, Phasing, and Refinement Statistics. (PDF 35 kb)

Supplementary Table 2

Mutational analysis of ESCRT-II assembly and function. (PDF 20 kb)

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Hierro, A., Sun, J., Rusnak, A. et al. Structure of the ESCRT-II endosomal trafficking complex. Nature 431, 221–225 (2004). https://doi.org/10.1038/nature02914

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