Functional and Physical Interaction Between Yeast Hsp90 and Hsp70

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 2018 Feb 22

PMID 29463764

Citations 44

Authors

Andrea N Kravats

Joel R Hoskins

Michael Reidy

Jill L Johnson

Shannon M Doyle

Olivier Genest

Daniel C Masison

Sue Wickner

Affiliations

Soon will be listed here.

Abstract

Heat shock protein 90 (Hsp90) is a highly conserved ATP-dependent molecular chaperone that is essential in eukaryotes. It is required for the activation and stabilization of more than 200 client proteins, including many kinases and steroid hormone receptors involved in cell-signaling pathways. Hsp90 chaperone activity requires collaboration with a subset of the many Hsp90 cochaperones, including the Hsp70 chaperone. In higher eukaryotes, the collaboration between Hsp90 and Hsp70 is indirect and involves Hop, a cochaperone that interacts with both Hsp90 and Hsp70. Here we show that yeast Hsp90 (Hsp82) and yeast Hsp70 (Ssa1), directly interact in vitro in the absence of the yeast Hop homolog (Sti1), and identify a region in the middle domain of yeast Hsp90 that is required for the interaction. In vivo results using Hsp90 substitution mutants showed that several residues in this region were important or essential for growth at high temperature. Moreover, mutants in this region were defective in interaction with Hsp70 in cell lysates. In vitro, the purified Hsp82 mutant proteins were defective in direct physical interaction with Ssa1 and in protein remodeling in collaboration with Ssa1 and cochaperones. This region of Hsp90 is also important for interactions with several Hsp90 cochaperones and client proteins, suggesting that collaboration between Hsp70 and Hsp90 in protein remodeling may be modulated through competition between Hsp70 and Hsp90 cochaperones for the interaction surface.

Citing Articles

Unveiling the thermotolerance mechanism of Pichia kudriavzevii LC375240 through transcriptomic and genetic analyses.

Qi Y, Qin Q, Ma J, Wang B, Jin C, Fang W BMC Biol. 2025; 23(1):55.

PMID: 39988652 PMC: 11849260. DOI: 10.1186/s12915-025-02159-1.

Hsp90, DnaK, and ClpB collaborate in protein reactivation.

Hoskins J, Wickramaratne A, Jewell C, Jenkins L, Wickner S Proc Natl Acad Sci U S A. 2025; 122(5):e2422640122.

PMID: 39879241 PMC: 11804706. DOI: 10.1073/pnas.2422640122.

Quantitative proteomic analysis reveals unique Hsp90 cycle-dependent client interactions.

Rios E, Goncalves D, Morano K, Johnson J Genetics. 2024; 227(2).

PMID: 38606935 PMC: 11151932. DOI: 10.1093/genetics/iyae057.

Hsp90, a team player in protein quality control and the stress response in bacteria.

Wickramaratne A, Wickner S, Kravats A Microbiol Mol Biol Rev. 2024; 88(2):e0017622.

PMID: 38534118 PMC: 11332350. DOI: 10.1128/mmbr.00176-22.

Insights into Hsp90 mechanism and functions learned from studies in the yeast, .

Rios E, Hunsberger I, Johnson J Front Mol Biosci. 2024; 11:1325590.

PMID: 38389899 PMC: 10881880. DOI: 10.3389/fmolb.2024.1325590.

References

Reidy M, Sharma R, Roberts B, Masison D . Human J-protein DnaJB6b Cures a Subset of Saccharomyces cerevisiae Prions and Selectively Blocks Assembly of Structurally Related Amyloids. J Biol Chem. 2015; 291(8):4035-47. PMC: 4759180. DOI: 10.1074/jbc.M115.700393. View

Flom G, Lemieszek M, Fortunato E, Johnson J . Farnesylation of Ydj1 is required for in vivo interaction with Hsp90 client proteins. Mol Biol Cell. 2008; 19(12):5249-58. PMC: 2592663. DOI: 10.1091/mbc.e08-04-0435. View

Bertelsen E, Chang L, Gestwicki J, Zuiderweg E . Solution conformation of wild-type E. coli Hsp70 (DnaK) chaperone complexed with ADP and substrate. Proc Natl Acad Sci U S A. 2009; 106(21):8471-6. PMC: 2689011. DOI: 10.1073/pnas.0903503106. View

Kravats A, Doyle S, Hoskins J, Genest O, Doody E, Wickner S . Interaction of E. coli Hsp90 with DnaK Involves the DnaJ Binding Region of DnaK. J Mol Biol. 2016; 429(6):858-872. PMC: 5357148. DOI: 10.1016/j.jmb.2016.12.014. View

Street T, Lavery L, Agard D . Substrate binding drives large-scale conformational changes in the Hsp90 molecular chaperone. Mol Cell. 2011; 42(1):96-105. PMC: 3105473. DOI: 10.1016/j.molcel.2011.01.029. View

Meyer P, Prodromou C, Liao C, Hu B, Roe S, Vaughan C . Structural basis for recruitment of the ATPase activator Aha1 to the Hsp90 chaperone machinery. EMBO J. 2004; 23(6):1402-10. PMC: 381413. DOI: 10.1038/sj.emboj.7600141. View

Zuehlke A, Reidy M, Lin C, LaPointe P, Alsomairy S, Lee D . An Hsp90 co-chaperone protein in yeast is functionally replaced by site-specific posttranslational modification in humans. Nat Commun. 2017; 8:15328. PMC: 5458067. DOI: 10.1038/ncomms15328. View

Genest O, Hoskins J, Camberg J, Doyle S, Wickner S . Heat shock protein 90 from Escherichia coli collaborates with the DnaK chaperone system in client protein remodeling. Proc Natl Acad Sci U S A. 2011; 108(20):8206-11. PMC: 3100916. DOI: 10.1073/pnas.1104703108. View

Prodromou C, Siligardi G, OBrien R, Woolfson D, Regan L, Panaretou B . Regulation of Hsp90 ATPase activity by tetratricopeptide repeat (TPR)-domain co-chaperones. EMBO J. 1999; 18(3):754-62. PMC: 1171168. DOI: 10.1093/emboj/18.3.754. View

10.

Morishima Y, Murphy P, Li D, Sanchez E, Pratt W . Stepwise assembly of a glucocorticoid receptor.hsp90 heterocomplex resolves two sequential ATP-dependent events involving first hsp70 and then hsp90 in opening of the steroid binding pocket. J Biol Chem. 2000; 275(24):18054-60. DOI: 10.1074/jbc.M000434200. View

11.

Genest O, Reidy M, Street T, Hoskins J, Camberg J, Agard D . Uncovering a region of heat shock protein 90 important for client binding in E. coli and chaperone function in yeast. Mol Cell. 2012; 49(3):464-73. PMC: 3570620. DOI: 10.1016/j.molcel.2012.11.017. View

12.

Sahasrabudhe P, Rohrberg J, Biebl M, Rutz D, Buchner J . The Plasticity of the Hsp90 Co-chaperone System. Mol Cell. 2017; 67(6):947-961.e5. DOI: 10.1016/j.molcel.2017.08.004. View

13.

Morgner N, Schmidt C, Beilsten-Edmands V, Ebong I, Patel N, Clerico E . Hsp70 forms antiparallel dimers stabilized by post-translational modifications to position clients for transfer to Hsp90. Cell Rep. 2015; 11(5):759-69. PMC: 4431665. DOI: 10.1016/j.celrep.2015.03.063. View

14.

Baindur-Hudson S, Edkins A, Blatch G . Hsp70/Hsp90 organising protein (hop): beyond interactions with chaperones and prion proteins. Subcell Biochem. 2014; 78:69-90. DOI: 10.1007/978-3-319-11731-7_3. View

15.

Johnson B, Chadli A, Felts S, Bouhouche I, Catelli M, Toft D . Hsp90 chaperone activity requires the full-length protein and interaction among its multiple domains. J Biol Chem. 2000; 275(42):32499-507. DOI: 10.1074/jbc.M005195200. View

16.

Nakamoto H, Fujita K, Ohtaki A, Watanabe S, Narumi S, Maruyama T . Physical interaction between bacterial heat shock protein (Hsp) 90 and Hsp70 chaperones mediates their cooperative action to refold denatured proteins. J Biol Chem. 2014; 289(9):6110-9. PMC: 3937677. DOI: 10.1074/jbc.M113.524801. View

17.

Gassler C, Buchberger A, Laufen T, Mayer M, Schroder H, Valencia A . Mutations in the DnaK chaperone affecting interaction with the DnaJ cochaperone. Proc Natl Acad Sci U S A. 1998; 95(26):15229-34. PMC: 28025. DOI: 10.1073/pnas.95.26.15229. View

18.

Hernandez M, Sullivan W, Toft D . The assembly and intermolecular properties of the hsp70-Hop-hsp90 molecular chaperone complex. J Biol Chem. 2002; 277(41):38294-304. DOI: 10.1074/jbc.M206566200. View

19.

Nathan D, Lindquist S . Mutational analysis of Hsp90 function: interactions with a steroid receptor and a protein kinase. Mol Cell Biol. 1995; 15(7):3917-25. PMC: 230631. DOI: 10.1128/MCB.15.7.3917. View

20.

Shiau A, Harris S, Southworth D, Agard D . Structural Analysis of E. coli hsp90 reveals dramatic nucleotide-dependent conformational rearrangements. Cell. 2006; 127(2):329-40. DOI: 10.1016/j.cell.2006.09.027. View