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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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.
The authors declare no competing financial interests.
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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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