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Asymmetric deceleration of ClpB or Hsp104 ATPase activity unleashes protein-remodeling activity

Nature Structural & Molecular Biology volume 14, pages 114–122 (2007)Cite this article

Abstract

Two members of the AAA+ superfamily, ClpB and Hsp104, collaborate with Hsp70 and Hsp40 to rescue aggregated proteins. However, the mechanisms that elicit and underlie their protein-remodeling activities remain unclear. We report that for both Hsp104 and ClpB, mixtures of ATP and ATP-γS unexpectedly unleash activation, disaggregation and unfolding activities independent of cochaperones. Mutations reveal how remodeling activities are elicited by impaired hydrolysis at individual nucleotide-binding domains. However, for some substrates, mixtures of ATP and ATP-γS abolish remodeling, whereas for others, ATP binding without hydrolysis is sufficient. Remodeling of different substrates necessitates a diverse balance of polypeptide 'holding' (which requires ATP binding but not hydrolysis) and unfolding (which requires ATP hydrolysis). We suggest that this versatility in reaction mechanism enables ClpB and Hsp104 to reactivate the entire aggregated proteome after stress and enables Hsp104 to control prion inheritance.

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Figure 1: Protein activation by ClpB or Hsp104 alone.
Figure 2: Reactivation of heat-aggregated proteins by ClpB or Hsp104 alone.
Figure 3: Protein unfolding by ClpB or Hsp104 alone.
Figure 4: Protein remodeling by Hsp104 NBD mutants.
Figure 5: Protein remodeling by ClpB NBD mutants.
Figure 6: Nucleotide hydrolysis by ClpB and Hsp104.
Figure 7: Effects of ATP and ATP-γS on prion remodeling by Hsp104.

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Acknowledgements

This research was supported by the Intramural Research Program of the NIH, National Cancer Institute, Center for Cancer Research, an American Heart Association scientist development grant to J.S and NIH grants to M.Z. (GM58626) and S.L. (GM25874). We thank C. Glabe (University of California, Irvine) for the antibody to oligomer and K. Mizuuchi and K. McKenney for helpful discussions.

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  1. Shannon M Doyle and James Shorter: These authors contributed equally to this work.

Authors and Affiliations

  1. Laboratory of Molecular Biology, National Cancer Institute, National Institutes of Health (NIH), Bethesda, 20892, Maryland, USA

    Shannon M Doyle, Joel R Hoskins & Sue Wickner

  2. Whitehead Institute for Biomedical Research, Nine Cambridge Center, Cambridge, 02142, Massachusetts, USA

    James Shorter & Susan Lindquist

  3. Department of Biochemistry, Kansas State University, Manhattan, 66506, Kansas, USA

    Michal Zolkiewski

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Contributions

S.M.D., J.S. and J.R.H. designed experiments, performed experiments, interpreted data and wrote the manuscript; M.Z., S.L. and S.W. designed experiments, interpreted data and wrote the manuscript.

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Correspondence to Susan Lindquist or Sue Wickner.

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Doyle, S., Shorter, J., Zolkiewski, M. et al. Asymmetric deceleration of ClpB or Hsp104 ATPase activity unleashes protein-remodeling activity. Nat Struct Mol Biol 14, 114–122 (2007). https://doi.org/10.1038/nsmb1198

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