Search tips
Search criteria 


Logo of jcinvestThe Journal of Clinical Investigation
J Clin Invest. 1991 February; 87(2): 680–686.
PMCID: PMC296359

Point mutation causing a single amino acid substitution in the hormone binding domain of the glucocorticoid receptor in familial glucocorticoid resistance.


Familial glucocorticoid resistance is a hypertensive, hyperandrogenic disorder characterized by increased serum cortisol concentrations in the absence of stigmata of Cushing's syndrome. Our previous studies of the first reported kindred showed a two- to threefold reduction in glucocorticoid receptor-ligand binding affinity in the propositus, and a lesser reduction in affinity in his mildly affected son and nephew. Glucocorticoid receptor cDNA from these three patients was amplified by polymerase chain reaction and sequenced. The cDNA nucleotide sequence was normal, except for nucleotide 2054, which substituted valine for aspartic acid at amino acid residue 641. The propositus was homozygous while the other relatives were heterozygous for the mutation. COS-7 monkey kidney cells were cotransfected with expression vectors for either wild type or Val 641-mutant receptors, together with the reporter plasmid pMMTV-CAT. Dexamethasone increased chloramphenicol acetyltransferase activity in cells expressing wild type receptor, but had no effect in cells expressing Val 641-mutant receptors, despite similar receptor concentrations, as indicated by Western blotting. The binding affinity for dexamethasone of the Val 641-mutant receptor was threefold lower than that of the wild type receptor. These results suggest that glucocorticoid resistance in this family is due to a point mutation in the steroid-binding domain of the glucocorticoid receptor.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.6M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Evans RM. The steroid and thyroid hormone receptor superfamily. Science. 1988 May 13;240(4854):889–895. [PubMed]
  • Hollenberg SM, Weinberger C, Ong ES, Cerelli G, Oro A, Lebo R, Thompson EB, Rosenfeld MG, Evans RM. Primary structure and expression of a functional human glucocorticoid receptor cDNA. Nature. 1985 Dec 19;318(6047):635–641. [PubMed]
  • Francke U, Foellmer BE. The glucocorticoid receptor gene is in 5q31-q32 [corrected]. Genomics. 1989 May;4(4):610–612. [PubMed]
  • Vingerhoeds AC, Thijssen JH, Schwarz F. Spontaneous hypercortisolism without Cushing's syndrome. J Clin Endocrinol Metab. 1976 Nov;43(5):1128–1133. [PubMed]
  • Chrousos GP, Vingerhoeds A, Brandon D, Eil C, Pugeat M, DeVroede M, Loriaux DL, Lipsett MB. Primary cortisol resistance in man. A glucocorticoid receptor-mediated disease. J Clin Invest. 1982 Jun;69(6):1261–1269. [PMC free article] [PubMed]
  • Linder MJ, Thompson EB. Abnormal glucocorticoid receptor gene and mRNA in primary cortisol resistance. J Steroid Biochem. 1989 Feb;32(2):243–249. [PubMed]
  • Lamberts SW, Poldermans D, Zweens M, de Jong FH. Familial cortisol resistance: differential diagnostic and therapeutic aspects. J Clin Endocrinol Metab. 1986 Dec;63(6):1328–1333. [PubMed]
  • Malchoff CD, Javier EC, Malchoff DM, Martin T, Rogol A, Brandon D, Loriaux DL, Reardon GE. Primary cortisol resistance presenting as isosexual precocity. J Clin Endocrinol Metab. 1990 Feb;70(2):503–507. [PubMed]
  • Chrousos GP, Vingerhoeds AC, Loriaux DL, Lipsett MB. Primary cortisol resistance: a family study. J Clin Endocrinol Metab. 1983 Jun;56(6):1243–1245. [PubMed]
  • Lipsett MB, Chrousos GP, Tomita M, Brandon DD, Loriaux DL. The defective glucocorticoid receptor in man and nonhuman primates. Recent Prog Horm Res. 1985;41:199–247. [PubMed]
  • Tomita M, Brandon DD, Chrousos GP, Vingerhoeds AC, Foster CM, Fowler D, Loriaux DL, Lipsett MB. Glucocorticoid receptors in Epstein-Barr virus-transformed lymphocytes from patients with glucocorticoid resistance and a glucocorticoid-resistant New World primate species. J Clin Endocrinol Metab. 1986 Jun;62(6):1145–1154. [PubMed]
  • Saiki RK, Gelfand DH, Stoffel S, Scharf SJ, Higuchi R, Horn GT, Mullis KB, Erlich HA. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. [PubMed]
  • Chomczynski P, Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. [PubMed]
  • Kadowaki T, Kadowaki H, Taylor SI. A nonsense mutation causing decreased levels of insulin receptor mRNA: detection by a simplified technique for direct sequencing of genomic DNA amplified by the polymerase chain reaction. Proc Natl Acad Sci U S A. 1990 Jan;87(2):658–662. [PubMed]
  • Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. [PubMed]
  • Southern EM. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. [PubMed]
  • Accili D, Frapier C, Mosthaf L, McKeon C, Elbein SC, Permutt MA, Ramos E, Lander E, Ullrich A, Taylor SI. A mutation in the insulin receptor gene that impairs transport of the receptor to the plasma membrane and causes insulin-resistant diabetes. EMBO J. 1989 Sep;8(9):2509–2517. [PubMed]
  • Ho SN, Hunt HD, Horton RM, Pullen JK, Pease LR. Site-directed mutagenesis by overlap extension using the polymerase chain reaction. Gene. 1989 Apr 15;77(1):51–59. [PubMed]
  • Giguère V, Hollenberg SM, Rosenfeld MG, Evans RM. Functional domains of the human glucocorticoid receptor. Cell. 1986 Aug 29;46(5):645–652. [PubMed]
  • Felgner PL, Gadek TR, Holm M, Roman R, Chan HW, Wenz M, Northrop JP, Ringold GM, Danielsen M. Lipofection: a highly efficient, lipid-mediated DNA-transfection procedure. Proc Natl Acad Sci U S A. 1987 Nov;84(21):7413–7417. [PubMed]
  • Gorman CM, Moffat LF, Howard BH. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol. 1982 Sep;2(9):1044–1051. [PMC free article] [PubMed]
  • Miesfeld R, Rusconi S, Godowski PJ, Maler BA, Okret S, Wikström AC, Gustafsson JA, Yamamoto KR. Genetic complementation of a glucocorticoid receptor deficiency by expression of cloned receptor cDNA. Cell. 1986 Aug 1;46(3):389–399. [PubMed]
  • Danielsen M, Northrop JP, Ringold GM. The mouse glucocorticoid receptor: mapping of functional domains by cloning, sequencing and expression of wild-type and mutant receptor proteins. EMBO J. 1986 Oct;5(10):2513–2522. [PubMed]
  • Arriza JL, Weinberger C, Cerelli G, Glaser TM, Handelin BL, Housman DE, Evans RM. Cloning of human mineralocorticoid receptor complementary DNA: structural and functional kinship with the glucocorticoid receptor. Science. 1987 Jul 17;237(4812):268–275. [PubMed]
  • Simons SS, Jr, Pumphrey JG, Rudikoff S, Eisen HJ. Identification of cysteine 656 as the amino acid of hepatoma tissue culture cell glucocorticoid receptors that is covalently labeled by dexamethasone 21-mesylate. J Biol Chem. 1987 Jul 15;262(20):9676–9680. [PubMed]
  • Miller NR, Simons SS., Jr Steroid binding to hepatoma tissue culture cell glucocorticoid receptors involves at least two sulfhydryl groups. J Biol Chem. 1988 Oct 15;263(29):15217–15225. [PubMed]
  • Beato M. Gene regulation by steroid hormones. Cell. 1989 Feb 10;56(3):335–344. [PubMed]
  • Simons SS, Jr, Miller PA, Wasner G, Miller NR, Mercier L. Inverse correlation between dexamethasone 21-mesylate agonist activity and sensitivity to dexamethasone for induction of tyrosine aminotransferase in rat hepatoma cells. J Steroid Biochem. 1988 Jul;31(1):1–7. [PubMed]
  • Brönnegård M, Werner S, Gustafsson JA. Primary cortisol resistance associated with a thermolabile glucocorticoid receptor in a patient with fatigue as the only symptom. J Clin Invest. 1986 Nov;78(5):1270–1278. [PMC free article] [PubMed]
  • Iida S, Gomi M, Moriwaki K, Itoh Y, Hirobe K, Matsuzawa Y, Katagiri S, Yonezawa T, Tarui S. Primary cortisol resistance accompanied by a reduction in glucocorticoid receptors in two members of the same family. J Clin Endocrinol Metab. 1985 May;60(5):967–971. [PubMed]
  • Nawata H, Sekiya K, Higuchi K, Kato K, Ibayashi H. Decreased deoxyribonucleic acid binding of glucocorticoid-receptor complex in cultured skin fibroblasts from a patient with the glucocorticoid resistance syndrome. J Clin Endocrinol Metab. 1987 Aug;65(2):219–226. [PubMed]
  • Griffin JE, Wilson JD. The syndromes of androgen resistance. N Engl J Med. 1980 Jan 24;302(4):198–209. [PubMed]
  • Marcelli M, Tilley WD, Wilson CM, Wilson JD, Griffin JE, McPhaul MJ. A single nucleotide substitution introduces a premature termination codon into the androgen receptor gene of a patient with receptor-negative androgen resistance. J Clin Invest. 1990 May;85(5):1522–1528. [PMC free article] [PubMed]
  • Hughes MR, Malloy PJ, Kieback DG, Kesterson RA, Pike JW, Feldman D, O'Malley BW. Point mutations in the human vitamin D receptor gene associated with hypocalcemic rickets. Science. 1988 Dec 23;242(4886):1702–1705. [PubMed]
  • Usala SJ, Tennyson GE, Bale AE, Lash RW, Gesundheit N, Wondisford FE, Accili D, Hauser P, Weintraub BD. A base mutation of the C-erbA beta thyroid hormone receptor in a kindred with generalized thyroid hormone resistance. Molecular heterogeneity in two other kindreds. J Clin Invest. 1990 Jan;85(1):93–100. [PMC free article] [PubMed]
  • Sakurai A, Takeda K, Ain K, Ceccarelli P, Nakai A, Seino S, Bell GI, Refetoff S, DeGroot LJ. Generalized resistance to thyroid hormone associated with a mutation in the ligand-binding domain of the human thyroid hormone receptor beta. Proc Natl Acad Sci U S A. 1989 Nov;86(22):8977–8981. [PubMed]

Articles from The Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation