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. 1993 Dec 1;90(23):11424–11428. doi: 10.1073/pnas.90.23.11424

Isolation of Hsp90 mutants by screening for decreased steroid receptor function.

S P Bohen 1, K R Yamamoto 1
PMCID: PMC47995  PMID: 8248264

Abstract

The 90-kDa heat shock protein Hsp90 represents a highly conserved strongly expressed gene family; in Saccharomyces cerevisiae, Hsp90 proteins are essential for cell viability. Hsp90 interacts with certain cellular proteins, including steroid hormone receptors, tyrosine and serine/threonine kinases, and other heat shock proteins, but its biological functions are not understood. The unliganded glucocorticoid receptor must interact with Hsp90 to acquire competence for high-affinity hormone binding and subsequent transcriptional regulation. By screening in yeast for defects in glucocorticoid receptor function, Hsp90 mutants were isolated. Four such mutants are described, all of which interact with the glucocorticoid receptor but display distinct defects in ligand responsiveness and differences in growth and resistance to high temperature.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bagchi M. K., Tsai S. Y., Tsai M. J., O'Malley B. W. Identification of a functional intermediate in receptor activation in progesterone-dependent cell-free transcription. Nature. 1990 Jun 7;345(6275):547–550. doi: 10.1038/345547a0. [DOI] [PubMed] [Google Scholar]
  2. Borkovich K. A., Farrelly F. W., Finkelstein D. B., Taulien J., Lindquist S. hsp82 is an essential protein that is required in higher concentrations for growth of cells at higher temperatures. Mol Cell Biol. 1989 Sep;9(9):3919–3930. doi: 10.1128/mcb.9.9.3919. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bresnick E. H., Dalman F. C., Sanchez E. R., Pratt W. B. Evidence that the 90-kDa heat shock protein is necessary for the steroid binding conformation of the L cell glucocorticoid receptor. J Biol Chem. 1989 Mar 25;264(9):4992–4997. [PubMed] [Google Scholar]
  4. Brugge J. S., Erikson E., Erikson R. L. The specific interaction of the Rous sarcoma virus transforming protein, pp60src, with two cellular proteins. Cell. 1981 Aug;25(2):363–372. doi: 10.1016/0092-8674(81)90055-6. [DOI] [PubMed] [Google Scholar]
  5. Brugge J., Yonemoto W., Darrow D. Interaction between the Rous sarcoma virus transforming protein and two cellular phosphoproteins: analysis of the turnover and distribution of this complex. Mol Cell Biol. 1983 Jan;3(1):9–19. doi: 10.1128/mcb.3.1.9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chakraborti P. K., Hoeck W., Groner B., Simons S. S., Jr Localization of the vicinal dithiols involved in steroid binding to the rat glucocorticoid receptor. Endocrinology. 1990 Nov;127(5):2530–2539. doi: 10.1210/endo-127-5-2530. [DOI] [PubMed] [Google Scholar]
  7. Chambraud B., Berry M., Redeuilh G., Chambon P., Baulieu E. E. Several regions of human estrogen receptor are involved in the formation of receptor-heat shock protein 90 complexes. J Biol Chem. 1990 Nov 25;265(33):20686–20691. [PubMed] [Google Scholar]
  8. Christianson T. W., Sikorski R. S., Dante M., Shero J. H., Hieter P. Multifunctional yeast high-copy-number shuttle vectors. Gene. 1992 Jan 2;110(1):119–122. doi: 10.1016/0378-1119(92)90454-w. [DOI] [PubMed] [Google Scholar]
  9. Farrelly F. W., Finkelstein D. B. Complete sequence of the heat shock-inducible HSP90 gene of Saccharomyces cerevisiae. J Biol Chem. 1984 May 10;259(9):5745–5751. [PubMed] [Google Scholar]
  10. Gametchu B., Harrison R. W. Characterization of a monoclonal antibody to the rat liver glucocorticoid receptor. Endocrinology. 1984 Jan;114(1):274–279. doi: 10.1210/endo-114-1-274. [DOI] [PubMed] [Google Scholar]
  11. Garabedian M. J., Yamamoto K. R. Genetic dissection of the signaling domain of a mammalian steroid receptor in yeast. Mol Biol Cell. 1992 Nov;3(11):1245–1257. doi: 10.1091/mbc.3.11.1245. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Godowski P. J., Rusconi S., Miesfeld R., Yamamoto K. R. Glucocorticoid receptor mutants that are constitutive activators of transcriptional enhancement. Nature. 1987 Jan 22;325(6102):365–368. doi: 10.1038/325365a0. [DOI] [PubMed] [Google Scholar]
  13. Howard K. J., Holley S. J., Yamamoto K. R., Distelhorst C. W. Mapping the HSP90 binding region of the glucocorticoid receptor. J Biol Chem. 1990 Jul 15;265(20):11928–11935. [PubMed] [Google Scholar]
  14. Joab I., Radanyi C., Renoir M., Buchou T., Catelli M. G., Binart N., Mester J., Baulieu E. E. Common non-hormone binding component in non-transformed chick oviduct receptors of four steroid hormones. 1984 Apr 26-May 2Nature. 308(5962):850–853. doi: 10.1038/308850a0. [DOI] [PubMed] [Google Scholar]
  15. Kaufmann S. H., Wright W. W., Okret S., Wikström A. C., Gustafsson J. A., Shaper N. L., Shaper J. H. Evidence that rodent epididymal sperm contain the Mr approximately 94,000 glucocorticoid receptor but lack the Mr approximately 90,000 heat shock protein. Endocrinology. 1992 May;130(5):3074–3084. doi: 10.1210/endo.130.5.1572314. [DOI] [PubMed] [Google Scholar]
  16. Lindquist S., Craig E. A. The heat-shock proteins. Annu Rev Genet. 1988;22:631–677. doi: 10.1146/annurev.ge.22.120188.003215. [DOI] [PubMed] [Google Scholar]
  17. Miyata Y., Yahara I. The 90-kDa heat shock protein, HSP90, binds and protects casein kinase II from self-aggregation and enhances its kinase activity. J Biol Chem. 1992 Apr 5;267(10):7042–7047. [PubMed] [Google Scholar]
  18. Nadeau K., Das A., Walsh C. T. Hsp90 chaperonins possess ATPase activity and bind heat shock transcription factors and peptidyl prolyl isomerases. J Biol Chem. 1993 Jan 15;268(2):1479–1487. [PubMed] [Google Scholar]
  19. Okret S., Wikström A. C., Wrange O., Andersson B., Gustafsson J. A. Monoclonal antibodies against the rat liver glucocorticoid receptor. Proc Natl Acad Sci U S A. 1984 Mar;81(6):1609–1613. doi: 10.1073/pnas.81.6.1609. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Perdew G. H. Association of the Ah receptor with the 90-kDa heat shock protein. J Biol Chem. 1988 Sep 25;263(27):13802–13805. [PubMed] [Google Scholar]
  21. Perdew G. H., Whitelaw M. L. Evidence that the 90-kDa heat shock protein (HSP90) exists in cytosol in heteromeric complexes containing HSP70 and three other proteins with Mr of 63,000, 56,000, and 50,000. J Biol Chem. 1991 Apr 15;266(11):6708–6713. [PubMed] [Google Scholar]
  22. Picard D., Khursheed B., Garabedian M. J., Fortin M. G., Lindquist S., Yamamoto K. R. Reduced levels of hsp90 compromise steroid receptor action in vivo. Nature. 1990 Nov 8;348(6297):166–168. doi: 10.1038/348166a0. [DOI] [PubMed] [Google Scholar]
  23. Picard D., Salser S. J., Yamamoto K. R. A movable and regulable inactivation function within the steroid binding domain of the glucocorticoid receptor. Cell. 1988 Sep 23;54(7):1073–1080. doi: 10.1016/0092-8674(88)90122-5. [DOI] [PubMed] [Google Scholar]
  24. Pongratz I., Mason G. G., Poellinger L. Dual roles of the 90-kDa heat shock protein hsp90 in modulating functional activities of the dioxin receptor. Evidence that the dioxin receptor functionally belongs to a subclass of nuclear receptors which require hsp90 both for ligand binding activity and repression of intrinsic DNA binding activity. J Biol Chem. 1992 Jul 5;267(19):13728–13734. [PubMed] [Google Scholar]
  25. Rafestin-Oblin M. E., Couette B., Radanyi C., Lombes M., Baulieu E. E. Mineralocorticosteroid receptor of the chick intestine. Oligomeric structure and transformation. J Biol Chem. 1989 Jun 5;264(16):9304–9309. [PubMed] [Google Scholar]
  26. Renoir J. M., Buchou T., Baulieu E. E. Involvement of a non-hormone-binding 90-kilodalton protein in the nontransformed 8S form of the rabbit uterus progesterone receptor. Biochemistry. 1986 Oct 21;25(21):6405–6413. doi: 10.1021/bi00369a010. [DOI] [PubMed] [Google Scholar]
  27. Rexin M., Busch W., Gehring U. Protein components of the nonactivated glucocorticoid receptor. J Biol Chem. 1991 Dec 25;266(36):24601–24605. [PubMed] [Google Scholar]
  28. Rose D. W., Welch W. J., Kramer G., Hardesty B. Possible involvement of the 90-kDa heat shock protein in the regulation of protein synthesis. J Biol Chem. 1989 Apr 15;264(11):6239–6244. [PubMed] [Google Scholar]
  29. Rusconi S., Yamamoto K. R. Functional dissection of the hormone and DNA binding activities of the glucocorticoid receptor. EMBO J. 1987 May;6(5):1309–1315. doi: 10.1002/j.1460-2075.1987.tb02369.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Schena M., Yamamoto K. R. Mammalian glucocorticoid receptor derivatives enhance transcription in yeast. Science. 1988 Aug 19;241(4868):965–967. doi: 10.1126/science.3043665. [DOI] [PubMed] [Google Scholar]
  31. Scherrer L. C., Hutchison K. A., Sanchez E. R., Randall S. K., Pratt W. B. A heat shock protein complex isolated from rabbit reticulocyte lysate can reconstitute a functional glucocorticoid receptor-Hsp90 complex. Biochemistry. 1992 Aug 18;31(32):7325–7329. doi: 10.1021/bi00147a017. [DOI] [PubMed] [Google Scholar]
  32. Schulman G., Bodine P. V., Litwack G. Modulators of the glucocorticoid receptor also regulate mineralocorticoid receptor function. Biochemistry. 1992 Feb 18;31(6):1734–1741. doi: 10.1021/bi00121a022. [DOI] [PubMed] [Google Scholar]
  33. Sikorski R. S., Hieter P. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics. 1989 May;122(1):19–27. doi: 10.1093/genetics/122.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Vegeto E., Allan G. F., Schrader W. T., Tsai M. J., McDonnell D. P., O'Malley B. W. The mechanism of RU486 antagonism is dependent on the conformation of the carboxy-terminal tail of the human progesterone receptor. Cell. 1992 May 15;69(4):703–713. doi: 10.1016/0092-8674(92)90234-4. [DOI] [PubMed] [Google Scholar]
  35. Wiech H., Buchner J., Zimmermann R., Jakob U. Hsp90 chaperones protein folding in vitro. Nature. 1992 Jul 9;358(6382):169–170. doi: 10.1038/358169a0. [DOI] [PubMed] [Google Scholar]
  36. Yamamoto K. R., Godowski P. J., Picard D. Ligand-regulated nonspecific inactivation of receptor function: a versatile mechanism for signal transduction. Cold Spring Harb Symp Quant Biol. 1988;53(Pt 2):803–811. doi: 10.1101/sqb.1988.053.01.091. [DOI] [PubMed] [Google Scholar]

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