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Cited by 57Cellular transport and secretionAutophagy in Disease and TherapyUbiquitin and proteasome pathways

A helical assembly of human ESCRT-I scaffolds reverse-topology membrane scission

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Flower, Thomas G., Takahashi, Yoshinori, Hudait, Arpa, Rose, Kevin, Tjahjono, Nicholas, Pak, Alexander J., Yokom, Adam L., Liang, Xinwen +4 more·2020·Nature Structural & Molecular Biology
DOI 10.1038/s41594-020-0426-4PMC PMC7339825

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Abstract

The ESCRT complexes drive membrane scission in HIV-1 release, autophagosome closure, multivesicular body biogenesis, cytokinesis, and other cell processes. ESCRT-I is the most upstream complex and bridges the system to HIV-1 Gag in virus release. The crystal structure of the headpiece of human ESCRT-I comprising TSG101-VPS28-VPS37B-MVB12A was determined, revealing an ESCRT-I helical assembly with a 12-molecule repeat. Electron microscopy confirmed that ESCRT-I subcomplexes form helical filaments in solution. Mutation of VPS28 helical interface residues blocks filament formation in vitro and autophagosome closure and HIV-1 release in human cells. Coarse-grained (CG) simulations of ESCRT assembly at HIV-1 budding sites suggest that formation of a 12-membered ring of ESCRT-I molecules is a geometry-dependent checkpoint during late stages of Gag assembly and HIV-1 budding and templates ESCRT-III assembly for membrane scission. These data show that ESCRT-I is not merely a bridging adaptor; it has an essential scaffolding and mechanical role in its own right.

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  • The functional human ESCRT-I complex is a hetero-tetramer of VPS28, TSG101, VPS37, and MVB12.

    “the functional ESCRT-I complex in humans organizes itself as a hetero-tetramer composed of a single copy of VPS28, TSG101, VPS37, and MVB12.”

    From Budding of a Retrovirus: Some Assemblies Required (2020) · cites this paper as [32]

    The paper’s introductory passage states that human ESCRT-I is a 1:1:1:1 complex containing TSG101, VPS28, a VPS37 protein, and a fourth subunit. MVB12A or MVB12B can fill that fourth position, but the passage also names UBAP1, UBA1L, and UMAD1; the citing sentence presents MVB12 as the sole option. (LLM, conf=0.86)

    Experimental — accuracy not yet measured

  • Both crystals are composed of helical assemblies of hetero-tetramer ESCRT-I complexes, and human VPS28-TSG101 subcomplex forms filaments in solution.

    “both crystals are composed of helical assemblies of the hetero-tetramer ESCRT-I complexes and a human VPS28-TSG101 subcomplex was shown to form filaments in solution as verified by negative stain electron microscopy [30,32].”

    From Budding of a Retrovirus: Some Assemblies Required (2020) · cites this paper as [32]

    The Results section describes helical assemblies in both the human and yeast crystals and reports that negative-stain electron microscopy identified human TSG101–VPS28 subcomplexes as filaments in solution. However, Extended Data Fig. 3 calls the helical yeast crystal assembly a trimeric ESCRT-I head, not a heterotetramer. Page numbers were not provided in our copy. (LLM, conf=0.90)

    Experimental — accuracy not yet measured

  • Human ESCRT-I complex solved via crystallography bore striking similarity to the homologous complex in yeast.

    “the human ESCRT-I complex was solved via crystallography and bore striking similarity to the homologous complex in yeast.”

    From Budding of a Retrovirus: Some Assemblies Required (2020) · cites this paper as [32]

    The Online methods (X-ray crystallography; page unavailable) describe solving a crystal structure of the human ESCRT-I head, not the entire complex. The Results (Helical assemblies; page unavailable) report remarkably similar human and yeast headpiece interfaces and tube dimensions, but also different helical arrangements. The claim is supported for the headpiece, while its reference to the human ESCRT-I complex as a whole is too broad. (LLM, conf=0.86)

    Experimental — accuracy not yet measured

  • The ESCRT-I complex has a globular head with descending alpha helical hairpins, VPS28 top, VPS37 bottom, and a coiled-coil stalk.

    “The complex assumes the structure of a globular head that is oriented as a descending stack of alpha helical hairpins with VPS28 on the top and VPS37 on the bottom, with a protruding coiled-coil stalk [29,30,31,32].”

    From Budding of a Retrovirus: Some Assemblies Required (2020) · cites this paper as [32]

    The paper’s background description calls the ESCRT-I headpiece fan-shaped, says it is capped by VPS28, and places it after a stalk. Its Methods place Vps37 at the base of a modeled yeast stalk, but our copy does not describe a globular head made of a descending stack of α-helical hairpins or establish that full arrangement for the human complex. (LLM, conf=0.82)

    Experimental — accuracy not yet measured

  • Contacts between neighboring VPS28 molecules are essential for higher-order assembly in yeast and human ESCRT-I filaments.

    “in both yeast and human ESCRT-I filaments, it appeared that contacts between neighboring VPS28 molecules were essential for higher-order assembly”

    From Budding of a Retrovirus: Some Assemblies Required (2020) · cites this paper as [32]

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Authors
Flower, T. G., Takahashi, Y., Hudait, A., Rose, K., Tjahjono, N., Pak, A. J., Yokom, A. L., Liang, X., Wang, H.-G., Bouamr, F., Voth, G. A., & Hurley, J. H.
Affiliations
  • QB3 (US)
  • Pennsylvania State University (US)
  • University of Chicago (US)
  • National Institutes of Health (US)
Corresponding author
Hurley, James H.
ORCID
Venue
Nature Structural & Molecular Biology
Publisher
Springer Science and Business Media LLC
Published
2020-05-18
Volume
27
Issue
6
Pages
570-580
ISSN
  • 1545-9985 (ISSN-L)
  • 1545-9993
DOI
10.1038/s41594-020-0426-4
PMC
PMC7339825
OpenAlex
W3024409383
Cited by
57
References
2
Citations by year
2026: 3 · 2025: 5 · 2024: 9 · 2023: 15 · 2022: 10 · 2021: 10 · 2020: 5
FWCI
4.99
Type
journal article
Language
en
Open access
Licence
Locations
  • Nature Structural & Molecular Biology (publishedVersion)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, public-domain)
  • PubMed (publishedVersion)
  • PubMed Central (acceptedVersion)
  • eScholarship (California Digital Library) (submittedVersion)
Funders
  • National Institutes of Health (GM127954)
  • National Cancer Institute (R01CA222349)
  • National Institute of General Medical Sciences (R01GM124149, R01GM128507, P30GM124169, R01GM127954)
  • National Institute of Allergy and Infectious Diseases (P50AI150464, F32AI150477, R01AI112442, R01AI150492, R37AI112442)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
  • Health Sciences › Medicine › Epidemiology › Autophagy in Disease and Therapy
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Ubiquitin and proteasome pathways
Keywords
ESCRT-I, membrane fission, helical assembly, HIV-1 budding, TSG101, VPS28, crystal structure, helical filaments, coarse-grained simulation, multivesicular body biogenesis, cytokinesis
MeSH terms
Autophagosomes, Tsg101 Protein, Cell Membrane, Crystallization, DNA-Binding Proteins, Humans, Phosphoproteins, Protein Conformation, Transcription Factors, HIV-1, Crystallography, X-Ray, Amino Acid Motifs, Adaptor Proteins, Signal Transducing, gag Gene Products, Human Immunodeficiency Virus, Host-Pathogen Interactions, Molecular Dynamics Simulation, Endosomal Sorting Complexes Required for Transport, Virus Release, HEK293 Cells

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