Evidence that the hormone-binding domain of the mouse glucocorticoid receptor directly represses DNA binding activity in a major portion of receptors that are "misfolded" after removal of hsp90 - PubMed (original) (raw)
. 1992 Feb 15;267(5):3190-5.
Affiliations
- PMID: 1737773
Free article
Evidence that the hormone-binding domain of the mouse glucocorticoid receptor directly represses DNA binding activity in a major portion of receptors that are "misfolded" after removal of hsp90
K A Hutchison et al. J Biol Chem. 1992.
Free article
Abstract
After dissociation of cytosolic heteromeric glucocorticoid receptor complexes by steroid, salt, and other methods, only 35-60% of the dissociated receptors can bind to DNA-cellulose. The DNA-binding and non-DNA-binding forms of the dissociated receptors have the same Mr and are phosphorylated to the same extent (Tienrungroj, W., Sanchez, E. R., Housley, P. R., Harrison, R. W., and Pratt, W. B. (1987) J. Biol. Chem. 262, 17347-17349). The basis for the different DNA-binding activities is unknown, but the DNA-binding fraction of the receptor has a more basic pI than the non-DNA-binding fraction (Smith, A. C., Elsasser, M. S., and Harmon, J. M. (1986) J. Biol. Chem. 261, 13285-13292). We have separated the non-DNA-binding state of the receptor from the DNA-binding state and then cleaved it with trypsin and chymotrypsin. We find that the 15-kDa tryptic fragment derived from the non-DNA-binding state of the dissociated receptor is fully competent in binding DNA, whereas the 42-kDa chymotryptic fragment containing both the hormone-binding and DNA-binding domains does not bind DNA. Trypsin cleavage of the molybdate-stabilized untransformed receptor also yields a 15-kDa fragment that is fully competent in binding DNA. Reducing agents do not restore DNA-binding to the non-DNA-binding fraction of the receptor and the hormone-binding domain can be separated from the DNA-binding domain on nonreducing gel electrophoresis. These results argue that the two domains are not linked by disulfide bridges, and they are consistent with the proposal that there are two least energy states of folding after dissociation of hsp90. A significant portion of the receptors is "misfolded" in such a manner that the steroid binding domain is directly preventing DNA-binding activity.
Similar articles
- Relationship of the 90-kDa murine heat shock protein to the untransformed and transformed states of the L cell glucocorticoid receptor.
Sanchez ER, Meshinchi S, Tienrungroj W, Schlesinger MJ, Toft DO, Pratt WB. Sanchez ER, et al. J Biol Chem. 1987 May 25;262(15):6986-91. J Biol Chem. 1987. PMID: 3294824 - Evidence that the 90-kDa phosphoprotein associated with the untransformed L-cell glucocorticoid receptor is a murine heat shock protein.
Sanchez ER, Toft DO, Schlesinger MJ, Pratt WB. Sanchez ER, et al. J Biol Chem. 1985 Oct 15;260(23):12398-401. J Biol Chem. 1985. PMID: 3900074 - The cyclosporin A-binding immunophilin CyP-40 and the FK506-binding immunophilin hsp56 bind to a common site on hsp90 and exist in independent cytosolic heterocomplexes with the untransformed glucocorticoid receptor.
Owens-Grillo JK, Hoffmann K, Hutchison KA, Yem AW, Deibel MR Jr, Handschumacher RE, Pratt WB. Owens-Grillo JK, et al. J Biol Chem. 1995 Sep 1;270(35):20479-84. doi: 10.1074/jbc.270.35.20479. J Biol Chem. 1995. PMID: 7657624 - Transformation of glucocorticoid and progesterone receptors to the DNA-binding state.
Pratt WB. Pratt WB. J Cell Biochem. 1987 Sep;35(1):51-68. doi: 10.1002/jcb.240350105. J Cell Biochem. 1987. PMID: 3312247 Review.
Cited by
- Breaking the mold: transcription factors in the anucleate platelet and platelet-derived microparticles.
Lannan KL, Sahler J, Kim N, Spinelli SL, Maggirwar SB, Garraud O, Cognasse F, Blumberg N, Phipps RP. Lannan KL, et al. Front Immunol. 2015 Feb 13;6:48. doi: 10.3389/fimmu.2015.00048. eCollection 2015. Front Immunol. 2015. PMID: 25762994 Free PMC article. Review. - Interaction of glucocorticosteroid receptor and wild-type or mutated 90-kDa heat shock protein coexpressed in baculovirus-infected Sf9 cells.
Cadepond F, Binart N, Chambraud B, Jibard N, Schweizer-Groyer G, Segard-Maurel I, Baulieu EE. Cadepond F, et al. Proc Natl Acad Sci U S A. 1993 Nov 15;90(22):10434-8. doi: 10.1073/pnas.90.22.10434. Proc Natl Acad Sci U S A. 1993. PMID: 8248127 Free PMC article. - Glucocorticoid receptor complexes form cooperatively with the Hsp90 co-chaperones Pp5 and FKBPs.
Kaziales A, Barkovits K, Marcus K, Richter K. Kaziales A, et al. Sci Rep. 2020 Jul 1;10(1):10733. doi: 10.1038/s41598-020-67645-8. Sci Rep. 2020. PMID: 32612187 Free PMC article. - The p23 molecular chaperones act at a late step in intracellular receptor action to differentially affect ligand efficacies.
Freeman BC, Felts SJ, Toft DO, Yamamoto KR. Freeman BC, et al. Genes Dev. 2000 Feb 15;14(4):422-34. Genes Dev. 2000. PMID: 10691735 Free PMC article. - Glucocorticoid receptor phosphorylation differentially affects target gene expression.
Chen W, Dang T, Blind RD, Wang Z, Cavasotto CN, Hittelman AB, Rogatsky I, Logan SK, Garabedian MJ. Chen W, et al. Mol Endocrinol. 2008 Aug;22(8):1754-66. doi: 10.1210/me.2007-0219. Epub 2008 May 15. Mol Endocrinol. 2008. PMID: 18483179 Free PMC article.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Research Materials
Miscellaneous