Abstract
Hsp90 (heat shock protein of 90 kDa) is a ubiquitous molecular chaperone responsible for the assembly and regulation of many eukaryotic signalling systems and is an emerging target for rational chemotherapy of many cancers. Although the structures of isolated domains of Hsp90 have been determined, the arrangement and ATP-dependent dynamics of these in the full Hsp90 dimer have been elusive and contentious. Here we present the crystal structure of full-length yeast Hsp90 in complex with an ATP analogue and the co-chaperone p23/Sba1. The structure reveals the complex architecture of the ‘closed’ state of the Hsp90 chaperone, the extensive interactions between domains and between protein chains, the detailed conformational changes in the amino-terminal domain that accompany ATP binding, and the structural basis for stabilization of the closed state by p23/Sba1. Contrary to expectations, the closed Hsp90 would not enclose its client proteins but provides a bipartite binding surface whose formation and disruption are coupled to the chaperone ATPase cycle.
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References
Panaretou, B. et al. ATP binding and hydrolysis are essential to the function of the Hsp90 molecular chaperone in vivo. EMBO J. 17, 4829–4836 (1998)
Obermann, W. M. J., Sondermann, H., Russo, A. A., Pavletich, N. P. & Hartl, F. U. In vivo function of Hsp90 is dependent on ATP binding and ATP hydrolysis. J. Cell Biol. 143, 901–910 (1998)
Mimnaugh, E. G., Chavany, C. & Neckers, L. Polyubiquitination and proteasomal degradation of the p185c-erbB-2 receptor protein-tyrosine kinase induced by geldanamycin. J. Biol. Chem. 271, 22796–22801 (1996)
Schneider, C. et al. Pharmacologic shifting of a balance between protein folding and degradation mediated by Hsp90. Proc. Natl Acad. Sci. USA 93, 14536–14541 (1996)
Pearl, L. H. Hsp90 and Cdc37—a chaperone cancer conspiracy. Curr. Opin. Genet. Dev. 15, 55–61 (2005)
Workman, P. Combinatorial attack on multistep oncogenesis by inhibiting the Hsp90 molecular chaperone. Cancer Lett. 206, 149–157 (2004)
Panaretou, B. et al. Activation of the ATPase activity of Hsp90 by the stress-regulated co-chaperone Aha1. Mol. Cell 10, 1307–1318 (2002)
Prodromou, C. et al. Regulation of Hsp90 ATPase activity by tetratricopeptide repeat (TPR)-domain co-chaperones. EMBO J. 18, 754–762 (1999)
Siligardi, G. et al. Regulation of Hsp90 ATPase activity by the co-chaperone Cdc37p/p50cdc37. J. Biol. Chem. 277, 20151–20159 (2002)
Johnson, J. L. & Toft, D. O. Binding of p23 and hsp90 during assembly with the progesterone receptor. Mol. Endocrinol. 9, 670–678 (1995)
Fang, Y., Fliss, A. E., Rao, J. & Caplan, A. J. SBA1 encodes a yeast hsp90 cochaperone that is homologous to vertebrate p23 proteins. Mol. Cell. Biol. 18, 3727–3734 (1998)
Sullivan, W. et al. Nucleotides and two functional states of hsp90. J. Biol. Chem. 272, 8007–8012 (1997)
Siligardi, G. et al. Co-chaperone regulation of conformational switching in the Hsp90 ATPase cycle. J. Biol. Chem. 279, 51989–51998 (2004)
McLaughlin, S. H., Smith, H. W. & Jackson, S. E. Stimulation of the weak ATPase activity of human Hsp90 by a client protein. J. Mol. Biol. 315, 787–798 (2002)
Richter, K., Walter, S. & Buchner, J. The co-chaperone Sba1 connects the ATPase reaction of Hsp90 to the progression of the chaperone cycle. J. Mol. Biol. 342, 1403–1413 (2004)
Pratt, W. B. & Dittmar, K. D. Studies with purified chaperones advance the understanding of the mechanism of glucocorticoid receptor hsp90 heterocomplex assembly. Trends Endocrinol. Metab. 9, 244–252 (1998)
Young, J. C. & Hartl, F. U. Polypeptide release by Hsp90 involves ATP hydrolysis and is enhanced by the co-chaperone p23. EMBO J. 19, 5930–5940 (2000)
Chadli, A. et al. Dimerization and N-terminal domain proximity underlie the function of the molecular chaperone heat shock protein 90. Proc. Natl Acad. Sci. USA 97, 12524–12529 (2000)
Prodromou, C. et al. The ATPase cycle of Hsp90 drives a molecular ‘clamp’ via transient dimerization of the N-terminal domains. EMBO J. 19, 4383–4392 (2000)
Huai, Q. et al. Structures of the N-terminal and middle domains of E. coli Hsp90 and conformation changes upon ADP binding. Structure 13, 579–590 (2005)
Immormino, R. M. et al. Ligand-induced conformational shift in the N-terminal domain of GRP94, an Hsp90 chaperone. J. Biol. Chem. 279, 46162–46171 (2004)
McLaughlin, S. H., Ventouras, L. A., Lobbezoo, B. & Jackson, S. E. Independent ATPase activity of Hsp90 subunits creates a flexible assembly platform. J. Mol. Biol. 344, 813–826 (2004)
Louvion, J. F., Warth, R. & Picard, D. Two eukaryote-specific regions of Hsp82 are dispensable for its viability and signal transduction functions in yeast. Proc. Natl Acad. Sci. USA 93, 13937–13942 (1996)
Prodromou, C., Roe, S. M., Piper, P. W. & Pearl, L. H. A molecular clamp in the crystal structure of the N-terminal domain of the yeast Hsp90 chaperone. Nature Struct. Biol. 4, 477–482 (1997)
Meyer, P. et al. Structural and functional analysis of the middle segment of Hsp90: Implications for ATP hydrolysis and client-protein and co-chaperone interactions. Mol. Cell 11, 647–658 (2003)
Weaver, A. J., Sullivan, W. P., Felts, S. J., Owen, B. A. & Toft, D. O. Crystal structure and activity of human p23, a heat shock protein 90 co-chaperone. J. Biol. Chem. 275, 23045–23052 (2000)
Prodromou, C. et al. Identification and structural characterization of the ATP/ADP-binding site in the Hsp90 molecular chaperone. Cell 90, 65–75 (1997)
Stebbins, C. E. et al. Crystal structure of an Hsp90-geldanamycin complex: Targetting of a protein chaperone by an antitumor agent. Cell 89, 239–250 (1997)
Roe, S. M. et al. The structural basis for inhibition of the Hsp90 molecular chaperone, by the anti-tumour antibiotics radicicol and geldanamycin. J. Med. Chem. 42, 260–266 (1999)
Cheung, K. M. et al. The identification, synthesis, protein crystal structure and in vitro biochemical evaluation of a new 3,4-diarylpyrazole class of Hsp90 inhibitors. Bioorg. Med. Chem. Lett. 15, 3338–3343 (2005)
Wright, L. et al. Structure-activity relationships in purine-based inhibitor binding to HSP90 isoforms. Chem. Biol. 11, 775–785 (2004)
Harris, S. F., Shiau, A. K. & Agard, D. A. The crystal structure of the carboxy-terminal dimerization domain of htpG, the Escherichia coli Hsp90, reveals a potential substrate binding site. Structure 12, 1087–1097 (2004)
Scheufler, C. et al. Structure of TPR domain-peptide complexes: critical elements in the assembly of the Hsp70-Hsp90 multichaperone machine. Cell 101, 199–210 (2000)
Bohen, S. P. & Yamamoto, K. R. Isolation of Hsp90 mutants by screening for decreased steroid receptor function. Proc. Natl Acad. Sci. USA 90, 11424–11428 (1993)
Nathan, D. F. & Lindquist, S. Mutational analysis of Hsp90 function: interactions with a steroid receptor and a protein kinase. Mol. Cell. Biol. 15, 3917–3925 (1995)
Ban, C., Junop, M. & Yang, W. Transformation of MutL by ATP binding and hydrolysis: a switch in DNA mismatch repair. Cell 97, 85–97 (1999)
Wigley, D. B., Davies, G. J., Dodson, E. J., Maxwell, A. & Dodson, G. Crystal structure of an N-terminal fragment of the DNA gyrase B protein. Nature 351, 624–629 (1991)
Meyer, P. et al. Structural basis for recruitment of the ATPase activator Aha1 to the Hsp90 chaperone machinery. EMBO J. 23, 511–519 (2004)
Oxelmark, E. et al. Genetic dissection of p23, an Hsp90 cochaperone, reveals a distinct surface involved in estrogen receptor signaling. J. Biol. Chem. 278, 36547–36555 (2003)
Lotz, G. P., Lin, H., Harst, A. & Obermann, W. M. J. Aha1 binds to the middle domain of Hsp90, contributes to client protein activation, and stimulates the ATPase activity of the molecular chaperone. J. Biol. Chem. 278, 17228–17235 (2003)
Harst, A., Lin, H. & Obermann, W. M. Aha1 competes with Hop, p50 and p23 for binding to the molecular chaperone Hsp90 and contributes to kinase and hormone receptor activation. Biochem. J. 387, 789–796 (2005)
Zhang, W. et al. Biochemical and structural studies of the interaction of Cdc37 with Hsp90. J. Mol. Biol. 340, 891–907 (2004)
Richter, K., Reinstein, J. & Buchner, J. N-terminal residues regulate the catalytic efficiency of the Hsp90 ATPase cycle. J. Biol. Chem. 277, 44905–44910 (2002)
Roe, S. M. et al. The mechanism of Hsp90 regulation by the protein kinase-specific cochaperone p50cdc37. Cell 116, 87–98 (2004)
Acknowledgements
We thank M. Gold, P. Wan, A. Doré and M. Kilkenny for assistance with synchrotron data collection. This work was supported by a Programme Grant from The Wellcome Trust and infrastructural support for structural biology at The Institute of Cancer Research, from Cancer Research UK.
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Coordinates and structure factors for the Hsp90–p23/Sba1–AMP-PNP complex and M-C construct have been deposited in the Protein Data Bank with accession codes 2CGE and 2CG9. Reprints and permissions information is available at npg.nature.com/reprintsandpermissions. The authors declare no competing financial interests.
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Supplementary Methods
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Supplementary Table
Table of crystallographic statistics. (DOC 56 kb)
Supplementary Figure 1
In the absence of ATP, Hsp90 conformation is unconstrained save for constitutive dimerisation via the C-domains. (PDF 59 kb)
Supplementary Notes
This file contains details of the significant proteins used in this study. (DOC 27 kb)
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Ali, M., Roe, S., Vaughan, C. et al. Crystal structure of an Hsp90–nucleotide–p23/Sba1 closed chaperone complex. Nature 440, 1013–1017 (2006). https://doi.org/10.1038/nature04716
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DOI: https://doi.org/10.1038/nature04716
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