1. Biochemistry and Chemical Biology
  2. Structural Biology and Molecular Biophysics

Symmetry broken and rebroken during the ATP hydrolysis cycle of the mitochondrial Hsp90 TRAP1

  1. Daniel Elnatan
  2. Miguel Betegon
  3. Yanxin Liu
  4. Theresa Ramelot
  5. Michael A Kennedy
  6. David A Agard  Is a corresponding author
  1. Howard Hughes Medical Institute, University of California, San Francisco, United Kingdom
  2. Howard Hughes Medical Institute, University of California, San Francisco, United States
  3. Miami University, United States
https://doi.org/10.7554/eLife.25235
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  1. Daniel Elnatan
  2. Miguel Betegon
  3. Yanxin Liu
  4. Theresa Ramelot
  5. Michael A Kennedy
  6. David A Agard
(2017)
Symmetry broken and rebroken during the ATP hydrolysis cycle of the mitochondrial Hsp90 TRAP1
eLife 6:e25235.
https://doi.org/10.7554/eLife.25235
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Abstract

Hsp90 is a homodimeric ATP-dependent molecular chaperone that remodels its substrate "client" proteins, facilitating their folding and activating them for biological function. Despite decades of research, the mechanism connecting ATP hydrolysis and chaperone function remains elusive. Particularly puzzling has been the apparent lack of cooperativity in hydrolysis of the ATP in each protomer. A crystal structure of the mitochondrial Hsp90, TRAP1, revealed that the catalytically active state is closed in a highly-strained asymmetric conformation. This asymmetry, unobserved in other Hsp90 homologs, is due to buckling of one of the protomers and is most pronounced at the broadly conserved client-binding region. Here, we show that rather than being cooperative or independent, ATP hydrolysis on the two protomers is sequential and deterministic. Moreover, dimer asymmetry sets up differential hydrolysis rates for each protomer, such that the buckled conformation favors ATP hydrolysis. Remarkably, after the first hydrolysis, the dimer undergoes a flip in the asymmetry while remaining in a closed state for the second hydrolysis. From these results, we propose a model where direct coupling of ATP hydrolysis and conformational flipping rearranges client-binding sites, providing a paradigm of how energy from ATP hydrolysis can be used for client remodeling.

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Author details

  1. Daniel Elnatan

    Department of Biochemistry and Biophysics, Howard Hughes Medical Institute, University of California, San Francisco, San Francisco, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-8359-0522

  2. Miguel Betegon

    Department of Biochemistry and Biophysics, Howard Hughes Medical Institute, University of California, San Francisco, San Francisco, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-7625-6190

  3. Yanxin Liu

    Department of Biochemistry and Biophysics, Howard Hughes Medical Institute, University of California, San Francisco, San Francisco, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-2253-3698

  4. Theresa Ramelot

    Department of Chemistry and Biochemistry, Miami University, Oxford, United States
    Competing interests
    The authors declare that no competing interests exist.

  5. Michael A Kennedy

    Department of Chemistry and Biochemistry, Miami University, Oxford, United States
    Competing interests
    The authors declare that no competing interests exist.

  6. David A Agard

    Department of Biochemistry and Biophysics, Howard Hughes Medical Institute, University of California, San Francisco, San Francisco, United States
    For correspondence
    agard@msg.ucsf.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-3512-695X

Funding

Howard Hughes Medical Institute

  • Daniel Elnatan
  • Miguel Betegon
  • Yanxin Liu
  • David A Agard

Helen Hay Whitney Foundation

  • Yanxin Liu

National Institutes of Health (U54-GM094597)

  • Michael A Kennedy

National Institutes of Health (U01-GM098254)

  • David A Agard

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Copyright

© 2017, Elnatan 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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