Structural Basis of Protein Translocation by the Vps4-Vta1 AAA ATPase

Abstract
Data availability
Article and author information
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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

The following data sets were generated

1. Monroe N
2. Han H
3. Shen PS
4. Sundquist WI
5. Hill CP
(2017) Vps4-Vta1 complex
Publicly available at the RCSB Protein Data Bank (accession no: 5UIE).

http://www.rcsb.org/pdb/explore/explore.do?structureId=5UIE
1. Monroe N
2. Han H
3. Shen PS
4. Sundquist WI
5. Hill CP
(2017) Vps4-Vta1 complex
Publicly available at the EMBL-EBI Protein Data Bank (accession no: EMD-8549).

http://www.ebi.ac.uk/pdbe/entry/emdb/EMD-8549
1. Monroe N
2. Han H
3. Shen PS
4. Sundquist WI
5. Hill CP
(2017) Vps4-Vta1 complex_sharpened map
Publicly available at the EMBL-EBI Protein Data Bank (accession no: EMD-8550).

http://www.ebi.ac.uk/pdbe/entry/emdb/EMD-8550
1. Monroe N
2. Han H
3. Shen PS
4. Sundquist WI
5. Hill CP
(2017) Vps4-HCP hexamer
Publicly available at the EMBL-EBI Protein Data Bank (accession no: EMD-8551).

http://www.ebi.ac.uk/pdbe/entry/emdb/EMD-8551
1. Monroe N
2. Han H
3. Shen PS
4. Sundquist WI
5. Hill CP
(2017) Vps4-Vta1 complex_VSL_A
Publicly available at the EMBL-EBI Protein Data Bank (accession no: EMD-8552).

http://www.ebi.ac.uk/pdbe/entry/emdb/EMD-8552
1. Monroe N
2. Han H
3. Shen PS
4. Sundquist WI
5. Hill CP
(2017) Vps4-Vta1 complex_VSL_B
Publicly available at the EMBL-EBI Protein Data Bank (accession no: EMD-8553).

http://www.ebi.ac.uk/pdbe/entry/emdb/EMD-8553
1. Monroe N
2. Han H
3. Shen PS
4. Sundquist WI
5. Hill CP
(2017) Vps4-Vta1 complex_VSL_C
Publicly available at the EMBL-EBI Protein Data Bank (accession no: EMD-8554).

http://www.ebi.ac.uk/pdbe/entry/emdb/EMD-8554
1. Monroe N
2. Han H
3. Shen PS
4. Sundquist WI
5. Hill CP
(2017) Vps4-Vta1 complex_VSL_D
Publicly available at the EMBL-EBI Protein Data Bank (accession no: EMD-8555).

http://www.ebi.ac.uk/pdbe/entry/emdb/EMD-8555
1. Monroe N
2. Han H
3. Shen PS
4. Sundquist WI
5. Hill CP
(2017) Vps4-Vta1 complex_VSL_E
Publicly available at the EMBL-EBI Protein Data Bank (accession no: EMD-8556).

http://www.ebi.ac.uk/pdbe/entry/emdb/EMD-8556
1. Monroe N
2. Han H
3. Shen PS
4. Sundquist WI
5. Hill CP
(2017) Vps4-Vta1 complex_VSL_F
Publicly available at the EMBL-EBI Protein Data Bank (accession no: EMD-8557).

http://www.ebi.ac.uk/pdbe/entry/emdb/EMD-8557
1. Monroe N
2. Han H
3. Shen PS
4. Sundquist WI
5. Hill CP
(2017) Vps4-Vta1 complex, State 3 of subunitF
Publicly available at the EMBL-EBI Protein Data Bank (accession no: EMD-8570).

http://www.ebi.ac.uk/pdbe/entry/emdb/EMD-8570
1. Monroe N
2. Han H
3. Shen PS
4. Sundquist WI
5. Hill CP
(2017) Vps4-Vta1 complex, State 2 of subunitF
Publicly available at the EMBL-EBI Protein Data Bank (accession no: EMD-8571).

http://www.ebi.ac.uk/pdbe/entry/emdb/EMD-8571
1. Monroe N
2. Han H
3. Shen PS
4. Sundquist WI
5. Hill CP
(2017) Vps4-Vta1 complex, State 1 of subunitF
Publicly available at the EMBL-EBI Protein Data Bank (accession no: EMD-8572).

http://www.ebi.ac.uk/pdbe/entry/emdb/EMD-8572

Article and author information

Author details

Nicole Monroe

Department of Biochemistry, University of Utah School of Medicine, Salt Lake City, United States

Competing interests
No competing interests declared.

"This ORCID iD identifies the author of this article:" 0000-0001-7678-4997
Han Han

Department of Biochemistry, University of Utah School of Medicine, Salt Lake City, United States

Competing interests
No competing interests declared.
Peter S Shen

Department of Biochemistry, University of Utah School of Medicine, Salt Lake City, United States

For correspondence
peter.shen@biochem.utah.edu

Competing interests
No competing interests declared.
Wesley I Sundquist

Department of Biochemistry, University of Utah School of Medicine, Salt Lake City, United States

For correspondence
wes@biochem.utah.edu

Competing interests
Wesley I Sundquist, Reviewing editor, eLife.

"This ORCID iD identifies the author of this article:" 0000-0001-9988-6021
Christopher P Hill

Department of Biochemistry, University of Utah School of Medicine, Salt Lake City, United States

For correspondence
chris@biochem.utah.edu

Competing interests
No competing interests declared.

"This ORCID iD identifies the author of this article:" 0000-0001-6796-7740

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.

Copyright

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.