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The ATPase cycle of Hsp90 drives a molecular 'clamp' via transient dimerization of the N-terminal domains
journal contribution
posted on 2023-06-08, 14:43 authored by Chrisostomos ProdromouChrisostomos Prodromou, Barry Panaretou, Shahzad Chohan, Giuliano Siligardi, Ronan O'Brien, John E Ladbury, S Mark Roe, Peter W Piper, Laurence PearlLaurence PearlHow 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.
History
Publication status
- Published
Journal
EMBO JournalISSN
0261-4189Publisher
Nature Publishing GroupExternal DOI
Issue
16Volume
19Page range
4383-92Department affiliated with
- Biochemistry Publications
Full text available
- No
Peer reviewed?
- Yes
Legacy Posted Date
2015-02-24Usage metrics
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Adenosine Triphosphatases/*metabolismAdenosine Triphosphate/metabolismAdenylyl Imidodiphosphate/metabolismBacterial Proteins/metabolismCircular DichroismCross-Linking Reagents/pharmacologyDNA GyraseDNA TopoisomerasesType II/metabolismDimerization*Escherichia coli ProteinsFungal Proteins/metabolismHSP90 Heat-Shock Proteins/*chemistry/genetics/*metabolismKineticsModelsBiologicalMolecularMolecular Chaperones/metabolismMutagenesisSite-DirectedMutationMissensePhenotypePlasmids/metabolismProtein BindingProtein ConformationProtein StructureSecondaryTertiary*Saccharomyces cerevisiae ProteinsSpectrometryFluorescenceTemperatureTime Factors
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