Cited by 492Heat shock proteins researchProtein Structure and DynamicsATP Synthase and ATPases Research

The ATPase cycle of Hsp90 drives a molecular ‘clamp’ via transient dimerization of the N‐terminal domains

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Prodromou, Chrisostomos, Panaretou, Barry, Chohan, Shahzad, Siligardi, Giuliano, O'Brien, Ronan, Ladbury, John E., Roe, S.Mark, Piper, Peter W. +1 more·2000·The EMBO Journal

Abstract

How the ATPase activity of Heat shock protein 90 (Hsp90) is coupled to client protein activation remains obscure. Using truncation and missense mutants of Hsp90, we analysed the structural implications of its ATPase cycle. C-terminal truncation mutants lacking inherent dimerization displayed reduced ATPase activity, but dimerized in the presence of 5'-adenylamido-diphosphate (AMP-PNP), and AMP-PNP- promoted association of N-termini in intact Hsp90 dimers was demonstrated. Recruitment of p23/Sba1 to C-terminal truncation mutants also required AMP-PNP-dependent dimerization. The temperature- sensitive (ts) mutant T101I had normal ATP affinity but reduced ATPase activity and AMP-PNP-dependent N-terminal association, whereas the ts mutant T22I displayed enhanced ATPase activity and AMP-PNP-dependent N-terminal dimerization, indicating a close correlation between these properties. The locations of these residues suggest that the conformation of the 'lid' segment (residues 100-121) couples ATP binding to N-terminal association. Consistent with this, a mutation designed to favour 'lid' closure (A107N) substantially enhanced ATPase activity and N-terminal dimerization. These data show that Hsp90 has a molecular 'clamp' mechanism, similar to DNA gyrase and MutL, whose opening and closing by transient N-terminal dimerization are directly coupled to the ATPase cycle.

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Crossref · 10.1093/emboj/19.16.4383

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Authors
Prodromou, C., Panaretou, B., Chohan, S., Siligardi, G., O'Brien, R., Ladbury, J. E., Roe, S. M., Piper, P. W., & Pearl, L. H.
Affiliations
  • Institute of Cancer Research (GB)
  • Imperial College London (GB)
  • King's College London (GB)
  • University College London (GB)
Corresponding author
Pearl, Laurence H.
Venue
The EMBO Journal
Published
2000-08-15
Volume
19
Issue
16
Pages
4383-4392
ISSN
  • 0261-4189 (ISSN-L)
  • 1460-2075
OpenAlex
W2046044279
Cited by
492
References
39
Citations by year
2026: 18 · 2025: 15 · 2024: 12 · 2023: 16 · 2022: 16 · 2021: 15 · 2020: 18 · 2019: 14 · 2018: 12 · 2017: 17 · 2016: 20 · 2015: 14 · 2014: 29 · 2013: 22 · 2012: 20
FWCI
6.14
Type
journal article
Language
en
Open access
Locations
  • The EMBO Journal (publishedVersion)
  • Europe PMC (PubMed Central) (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
  • UCL Discovery (University College London) (submittedVersion)
  • Figshare (submittedVersion, cc-by)
  • Research Portal (King's College London) (submittedVersion)
Funders
Wellcome Trust
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Heat shock proteins research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › ATP Synthase and ATPases Research
Keywords
HSP90, ATPase cycle, molecular clamp, AMP-PNP, temperature-sensitive mutants, ATP binding, protein dimerization, DNA gyrase, MutL
MeSH terms
MutL Proteins, Adenosine Triphosphatases, Adenosine Triphosphate, Adenylyl Imidodiphosphate, Bacterial Proteins, Circular Dichroism, Cross-Linking Reagents, DNA Topoisomerases, Type II, Fungal Proteins, Kinetics, Models, Biological, Models, Molecular, Phenotype, Plasmids, Protein Binding, Protein Conformation, Spectrometry, Fluorescence, Temperature, Time Factors, Mutagenesis, Site-Directed, Protein Structure, Secondary, Protein Structure, Tertiary, Molecular Chaperones, HSP90 Heat-Shock Proteins, Dimerization, Mutation, Missense, DNA Gyrase, Saccharomyces cerevisiae Proteins, Escherichia coli Proteins

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