Structural Basis of Protein Translocation by the Vps4-Vta1 AAA ATPase
Abstract
Many important cellular membrane fission reactions are driven by ESCRT pathways, which culminate in disassembly of ESCRT-III polymers by the AAA ATPase Vps4. We report a 4.3 Å resolution cryo-EM structure of the active Vps4 hexamer with its cofactor Vta1, ADP•BeFx, and an ESCRT-III substrate peptide. Four Vps4 subunits form a helix whose interfaces are consistent with ATP-binding, is stabilized by Vta1, and binds the substrate peptide. The fifth subunit approximately continues this helix but appears to be dissociating. The final Vps4 subunit completes a notched-washer configuration as if transitioning between the ends of the helix. We propose that ATP binding propagates growth at one end of the helix while hydrolysis promotes disassembly at the other end, so that Vps4 'walks' along ESCRT-III until it encounters the ordered N-terminal domain to destabilize the ESCRT-III lattice. This model may be generally applicable to other protein-translocating AAA ATPases.
Data availability
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Vps4-Vta1 complexPublicly available at the RCSB Protein Data Bank (accession no: 5UIE).
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Vps4-Vta1 complexPublicly available at the EMBL-EBI Protein Data Bank (accession no: EMD-8549).
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Vps4-Vta1 complex_sharpened mapPublicly available at the EMBL-EBI Protein Data Bank (accession no: EMD-8550).
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Vps4-HCP hexamerPublicly available at the EMBL-EBI Protein Data Bank (accession no: EMD-8551).
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Vps4-Vta1 complex_VSL_APublicly available at the EMBL-EBI Protein Data Bank (accession no: EMD-8552).
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Vps4-Vta1 complex_VSL_BPublicly available at the EMBL-EBI Protein Data Bank (accession no: EMD-8553).
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Vps4-Vta1 complex_VSL_CPublicly available at the EMBL-EBI Protein Data Bank (accession no: EMD-8554).
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Vps4-Vta1 complex_VSL_DPublicly available at the EMBL-EBI Protein Data Bank (accession no: EMD-8555).
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Vps4-Vta1 complex_VSL_EPublicly available at the EMBL-EBI Protein Data Bank (accession no: EMD-8556).
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Vps4-Vta1 complex_VSL_FPublicly available at the EMBL-EBI Protein Data Bank (accession no: EMD-8557).
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Vps4-Vta1 complex, State 3 of subunitFPublicly available at the EMBL-EBI Protein Data Bank (accession no: EMD-8570).
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Vps4-Vta1 complex, State 2 of subunitFPublicly available at the EMBL-EBI Protein Data Bank (accession no: EMD-8571).
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Vps4-Vta1 complex, State 1 of subunitFPublicly available at the EMBL-EBI Protein Data Bank (accession no: EMD-8572).
Article and author information
Author details
Funding
National Institutes of Health (P50 GM082545)
- Nicole Monroe
- Han Han
- Peter S Shen
- Wesley I Sundquist
- Christopher P Hill
National Institutes of Health (Microbial Pathogenesis Training Grant T32 AI055434)
- Nicole Monroe
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Reviewing Editor
- Sriram Subramaniam, National Cancer Institute, United States
Version history
- Received: December 21, 2016
- Accepted: April 4, 2017
- Accepted Manuscript published: April 5, 2017 (version 1)
- Version of Record published: May 2, 2017 (version 2)
Copyright
© 2017, Monroe et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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