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Molecular mechanism of multivesicular body biogenesis by ESCRT complexes

Nature volume 464, pages 864–869 (2010)Cite this article

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Abstract

When internalized receptors and other cargo are destined for lysosomal degradation, they are ubiquitinated and sorted by the endosomal sorting complex required for transport (ESCRT) complexes 0, I, II and III into multivesicular bodies. Multivesicular bodies are formed when cargo-rich patches of the limiting membrane of endosomes bud inwards by an unknown mechanism and are then cleaved to yield cargo-bearing intralumenal vesicles. The biogenesis of multivesicular bodies was reconstituted and visualized using giant unilamellar vesicles, fluorescent ESCRT-0, -I, -II and -III complexes, and a membrane-tethered fluorescent ubiquitin fusion as a model cargo. Here we show that ESCRT-0 forms domains of clustered cargo but does not deform membranes. ESCRT-I and ESCRT-II in combination deform the membrane into buds, in which cargo is confined. ESCRT-I and ESCRT-II localize to the bud necks, and recruit ESCRT-0-ubiquitin domains to the buds. ESCRT-III subunits localize to the bud neck and efficiently cleave the buds to form intralumenal vesicles. Intralumenal vesicles produced in this reaction contain the model cargo but are devoid of ESCRTs. The observations explain how the ESCRTs direct membrane budding and scission from the cytoplasmic side of the bud without being consumed in the reaction.

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Figure 1: Model cargo clustering by ESCRT-0.
Figure 2: ESCRT-I and ESCRT-II induce membrane buds and confine cargo in them.
Figure 3: ESCRT-I and ESCRT-II localize to the necks of membrane buds.
Figure 4: ESCRT-0 Ub domains co-localize with ESCRT-I–II membrane buds.
Figure 5: ESCRT-III localizes to bud necks for membrane scission.
Figure 6: Molecular mechanism of MVB biogenesis.

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Acknowledgements

We thank E. Boura for producing the CFP–Ub and GFP–Ub, J. Lippincott-Schwartz for microscope access, B. Wendland for a yeast strain and advice, and W. Prinz for comments on the manuscript. This work was funded by the Intramural Program of the National Institutes of Health, National Institute of Diabetes and Digestive and Kidney Diseases and Intramural AIDS Targeted Antiviral Program to J.H.H. and an EMBO long-term fellowship to T.W.

Author Contributions T.W. carried out all experiments; T.W. and J.H.H. designed experiments and analysed data; and J.H.H. wrote the manuscript.

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

    Thomas Wollert & James H. Hurley

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Correspondence to James H. Hurley.

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Wollert, T., Hurley, J. Molecular mechanism of multivesicular body biogenesis by ESCRT complexes. Nature 464, 864–869 (2010). https://doi.org/10.1038/nature08849

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