GR chaperone cycle mechanism revealed by cryo-EM: reactivation of GR by the GR:Hsp90:p23 client-maturation complex
Open accessPreprint — published as 10.1038/s41586-021-04236-1 · Paper page
Abstract
Hsp90 is a conserved and essential molecular chaperone responsible for the folding and activation of hundreds of ‘client’ proteins 1,2 . The glucocorticoid receptor (GR) is a model client that constantly depends on Hsp90 for activity 3 . Previously, we revealed GR ligand binding is inhibited by Hsp70 and restored by Hsp90, aided by the cochaperone p23 4 . However, a molecular understanding of the chaperone-induced transformations that occur between the inactive Hsp70:Hsp90 ‘client-loading complex’ and an activated Hsp90:p23 ‘client-maturation complex’ is lacking for GR, or for any client. Here, we present a 2.56Å cryo-EM structure of the GR-maturation complex (GR:Hsp90:p23), revealing that the GR ligand binding domain is, surprisingly, restored to a folded, ligand-bound conformation, while simultaneously threaded through the Hsp90 lumen. Also, unexpectedly, p23 directly stabilizes native GR using a previously uncharacterized C-terminal helix, resulting in enhanced ligand-binding. This is the highest resolution Hsp90 structure to date and the first atomic resolution structure of a client bound to Hsp90 in a native conformation, sharply contrasting with the unfolded kinase:Hsp90 structure 5 . Thus, aided by direct cochaperone:client interactions, Hsp90 dictates client-specific folding outcomes. Together with the GR-loading complex structure (Wang et al. 2020), we present the molecular mechanism of chaperone-mediated GR remodeling, establishing the first complete chaperone cycle for any client.
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Cryo-EM structures of client-bound Hsp90 show the client unfolds substantially, probably when the closed state forms.
“Cryo-EM structures of the client-bound Hsp90 reveal the client undergoes substantial unfolding, likely upon formation of the Hsp90 closed state”
From The structure of an Hsp90-immunophilin complex reveals cochaperone recognition of the client maturation state (2021) · cites this paper as Noddings et al., 2020
The Discussion (page unavailable) says GR is partially unfolded in the loading complex but becomes folded as Hsp90 reaches its fully closed maturation state—the opposite of unfolding upon closure. It also notes an unfolded kinase in another closed Hsp90 complex, so the structures do not establish unfolding as a general consequence of closure. (LLM, conf=0.88)
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Experimental — accuracy not yet measuredIn the GR:Hsp90:p23 structure p23 contacts the GR client directly and boosts ligand binding through its C-terminal tail helix, which is not resolved in the present structures.
“In the GR:Hsp90:p23 structure (Noddings et al., 2020), p23 directly contacts the GR client and enhances ligand binding via its C-terminal tail helix, which is unresolved in our structures.”
From The structure of an Hsp90-immunophilin complex reveals cochaperone recognition of the client maturation state (2021) · cites this paper as Noddings et al., 2020
In contrast to the yeast Hsp90:p23 crystal structure, the structures reported here and the GR:Hsp90:p23 cryo-EM structure indicate that one p23 molecule binds the human Hsp90 dimer in the maturation state.
“unlike the crystal structure of yeast Hsp90:p23, the structures presented here (Hsp90:FKBP51:23 and Hsp90:p23) as well as the recent cryo-EM structure of GR:Hsp90:p23 (Noddings et al., 2020), indicate a single p23 molecule appears to bind the human Hsp90 dimer in the maturation state”
From The structure of an Hsp90-immunophilin complex reveals cochaperone recognition of the client maturation state (2021) · cites this paper as Noddings et al., 2020
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Lee, K., Thwin, A. C., Nadel, C. M., Tse, E., Gates, S. N., Gestwicki, J. E., & Southworth, D. R. (2021) · Molecular Cell · doi:10.1016/j.molcel.2021.07.023 · Cited by 76 · see citing works
Biebl, M. M., Lopez, A., Rehn, A., Freiburger, L., Lawatscheck, J., Blank, B., Sattler, M., & Buchner, J. (2021) · Nature Communications · doi:10.1038/s41467-021-21063-0 · Cited by 58 · see citing works
Client binding shifts the populations of dynamic Hsp90 conformations through an allosteric network
Lopez, A., Dahiya, V., Delhommel, F., Freiburger, L., Stehle, R., Asami, S., Rutz, D., Blair, L., Buchner, J., & Sattler, M. (2021) · Science Advances · doi:10.1126/sciadv.abl7295 · Cited by 59 · see citing works
Cryo-EM reveals the dynamic interplay between mitochondrial Hsp90 and SdhB folding intermediates
Liu, Y., Elnatan, D., Sun, M., Myasnikov, A. G., & Agard, D. A. (2020) · doi:10.1101/2020.10.06.327627 · Cited by 16 · see citing works
Self-citation
- Authors
- Noddings, C. M., Wang, R. Y.-R., & Agard, D. A.
- Affiliations
- University of California, San Francisco (US)
- Corresponding author
- Agard, David A.
- Venue
- Nature
- Publisher
- openRxiv
- Published
- 2020-09-13
- Volume
- 601
- Issue
- 7893
- Pages
- 465-469
- ISSN
- 0028-0836 (ISSN-L)
- PMC
- PMC8994517
- OpenAlex
- W4200236070
- Cited by
- 166
- References
- 0
- Citations by year
- 2026: 30 · 2025: 23 · 2024: 35 · 2023: 34 · 2022: 32 · 2021: 9 · 2020: 2
- FWCI
- 8.71
- Type
- journal article
- Language
- en
- Open access
- green
- doi.org
- Licence
- Locations
- Nature (publishedVersion)
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
- Research Square (acceptedVersion, cc-by)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- RePEc: Research Papers in Economics (submittedVersion)
- eScholarship (California Digital Library) (submittedVersion)
- Funders
- Howard Hughes Medical Institute
- W. M. Keck Foundation
- Life Sciences Research Foundation
- National Institutes of Health (1s10od021741-01, 1s10od020054-01)
- University of California, San Francisco (R35GM118099)
- National Cancer Institute (F31CA265084)
- National Institute of General Medical Sciences (R35GM118099, P20GM104420, R01GM127675)
- 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 › Genomics and Chromatin Dynamics
- Keywords
- HSP90, p23, glucocorticoid receptor, molecular chaperones, ligand-binding domain, chaperone cycle, protein folding, co-chaperones, HSP70
- MeSH terms
- Prostaglandin-E Synthases, Humans, Ligands, Protein Binding, Receptors, Glucocorticoid, Molecular Chaperones, HSP70 Heat-Shock Proteins, HSP90 Heat-Shock Proteins, Cryoelectron Microscopy
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