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. 1985 Apr;75(4):1291–1296. doi: 10.1172/JCI111829

Reduced affinity of the androgen receptor for 5 alpha-dihydrotestosterone but not methyltrienolone in a form of partial androgen resistance. Studies on cultured genital skin fibroblasts.

L Pinsky, M Kaufman, A E Chudley
PMCID: PMC425458  PMID: 3872888

Abstract

We have studied a child with posterior labial fusion, clitoral phallus, female urethra, and a short, blind vagina born to a mother with decreased axillary and pubic hair. Her karyotype is 46,XY. At 2 yr of age, the child's basal level of plasma testosterone was less than 0.35 nM and after human chorionic gonadotropin stimulation, it rose to 2.6. Testis and epididymis histology were normal. Her cultured genital (labial) skin fibroblasts have normal testosterone 5 alpha-reductase activity, and metabolize 5 alpha-dihydrotestosterone (DHT) normally, but they do not augment (up-regulate) their basal androgen-receptor binding activity during prolonged incubation with DHT. With DHT, the androgen receptor in her genital skin fibroblasts has a normal binding capacity (maximum binding capacity = 25 fmol/mg protein), but an increased rate constant of dissociation (k = 11.6 X 10(-3) min-1; normal, 6 +/- 1.2 (+/- SD)), and a decreased apparent equilibrium binding affinity (Kd = 0.6 nM; normal, 0.22 +/- 0.09) that is evident in the results of 2-h assays but not of those lasting 0.5 h. With the synthetic androgen, methyltrienolone, all three binding properties of the receptor are normal, and her receptor activity up-regulates normally. We interpret these results to mean that the subject has a ligand-selective defect in the time-dependent transformation of initial, low-affinity androgen-receptor complexes to serial states of higher affinity, presumably as the result of a structural mutation at the X-linked locus that encodes the androgen receptor protein.

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Selected References

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  1. Aiman J., Griffin J. E. The frequency of androgen receptor deficiency in infertile men. J Clin Endocrinol Metab. 1982 Apr;54(4):725–732. doi: 10.1210/jcem-54-4-725. [DOI] [PubMed] [Google Scholar]
  2. Bonne C., Raynaud J. P. Methyltrienolone, a specific ligand for cellular androgen receptors. Steroids. 1975 Aug;26(2):227–232. doi: 10.1016/s0039-128x(75)80023-7. [DOI] [PubMed] [Google Scholar]
  3. Brown T. R., Maes M., Rothwell S. W., Migeon C. J. Human complete androgen insensitivity with normal dihydrotestosterone receptor binding capacity in cultured genital skin fibroblasts: evidence for a qualitative abnormality of the receptor. J Clin Endocrinol Metab. 1982 Jul;55(1):61–69. doi: 10.1210/jcem-55-1-61. [DOI] [PubMed] [Google Scholar]
  4. Brown T. R., Migeon C. J. Cultured human skin fibroblasts: a model for the study of androgen action. Mol Cell Biochem. 1981 Apr 13;36(1):3–22. doi: 10.1007/BF02354827. [DOI] [PubMed] [Google Scholar]
  5. Brown T. R., Rothwell S. W., Migeon C. J. Comparison of methyltrienolone and dihydrotestosterone binding and metabolism in human genital skin fibroblasts. J Steroid Biochem. 1981 Oct;14(10):1013–1022. doi: 10.1016/0022-4731(81)90209-0. [DOI] [PubMed] [Google Scholar]
  6. Eil C. Familial incomplete male pseudohermaphroditism associated with impaired nuclear androgen retention. Studies in cultured skin fibroblasts. J Clin Invest. 1983 Apr;71(4):850–858. doi: 10.1172/JCI110839. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Elawady M. K., Allman D. R., Griffin J. E., Wilson J. D. Expression of a mutant androgen receptor in cloned fibroblasts derived from a heterozygous carrier for the syndrome of testicular feminization. Am J Hum Genet. 1983 May;35(3):376–384. [PMC free article] [PubMed] [Google Scholar]
  8. FEYEL-CABANES T. ETUDE BIOLOGIQUE D'UN ANALOGUE TRI'ENIQUE DE LA M'ETHYLTEST'ERONE. C R Seances Soc Biol Fil. 1963 Dec 15;157:1428–1429. [PubMed] [Google Scholar]
  9. Griffin J. E., Durrant J. L. Qualitative receptor defects in families with androgen resistance: failure of stabilization of the fibroblast cytosol androgen receptor. J Clin Endocrinol Metab. 1982 Sep;55(3):465–474. doi: 10.1210/jcem-55-3-465. [DOI] [PubMed] [Google Scholar]
  10. Groner B., Ponta H., Rahmsdorf U., Herrlich P., Pfahl M., Hynes N. E. Glucocorticoid hormone interactions with cloned proviral DNA of mouse mammary tumor virus. J Steroid Biochem. 1984 Jan;20(1):95–98. doi: 10.1016/0022-4731(84)90194-8. [DOI] [PubMed] [Google Scholar]
  11. Hodgins M. B. Binding of androgens in 5 alpha-reductase-deficient human genital skin fibroblasts: inhibition by progesterone and its metabolites. J Endocrinol. 1982 Sep;94(3):415–427. doi: 10.1677/joe.0.0940415. [DOI] [PubMed] [Google Scholar]
  12. Jukier L., Kaufman M., Pinsky L., Peterson R. E. Partial androgen resistance associated with secondary 5 alpha-reductase deficiency: identification of a novel qualitative androgen receptor defect and clinical implications. J Clin Endocrinol Metab. 1984 Oct;59(4):679–688. doi: 10.1210/jcem-59-4-679. [DOI] [PubMed] [Google Scholar]
  13. Kaufman M., Pinsky L., Bowin A., Au M. W. Familial external genital ambiguity due to a transformation defect of androgen-receptor complexes that is expressed with 5 alpha-dihydrotestosterone and the synthetic androgen methyltrienolone. Am J Med Genet. 1984 Jul;18(3):493–507. doi: 10.1002/ajmg.1320180319. [DOI] [PubMed] [Google Scholar]
  14. Kaufman M., Pinsky L., Feder-Hollander R. Defective up-regulation of the androgen receptor in human androgen insensitivity. Nature. 1981 Oct 29;293(5835):735–737. doi: 10.1038/293735a0. [DOI] [PubMed] [Google Scholar]
  15. Kaufman M., Pinsky L., Hollander R., Bailey J. D. Regulation of the androgen receptor by androgen in normal and androgen-resistant genital skin fibroblasts. J Steroid Biochem. 1983 Apr;18(4):383–390. doi: 10.1016/0022-4731(83)90055-9. [DOI] [PubMed] [Google Scholar]
  16. Kaufman M., Pinsky L., Simard L., Wong S. C. Defective activation of androgen-receptor complexes: a marker of androgen insensitivity. Mol Cell Endocrinol. 1982 Feb;25(2):151–162. doi: 10.1016/0303-7207(82)90048-x. [DOI] [PubMed] [Google Scholar]
  17. Kaufman M., Straisfeld C., Pinsky L. Expression of androgen-responsive properties in human skin fibroblast strains of genital and nongenital origin. Somatic Cell Genet. 1977 Jan;3(1):17–25. doi: 10.1007/BF01550984. [DOI] [PubMed] [Google Scholar]
  18. Krüskemper H. L., Noell G. Liver toxicity of a new anabolic agent: methyltrienolone (17-alpha-methyl-4,9,11-estratriene-17 beta-ol-3-one). Steroids. 1966 Jul;8(1):13–24. doi: 10.1016/0039-128x(66)90114-0. [DOI] [PubMed] [Google Scholar]
  19. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  20. Larrea F., Benavides G., Scaglia H., Kofman-Alfaro S., Ferrusca E., Medina M., Pérez-Palacios G. Gynecomastia as a familial incomplete male pseudohermaphroditism type 1: a limited androgen resistance syndrome. J Clin Endocrinol Metab. 1978 Jun;46(6):961–970. doi: 10.1210/jcem-46-6-961. [DOI] [PubMed] [Google Scholar]
  21. Migeon B. R., Brown T. R., Axelman J., Migeon C. J. Studies of the locus for androgen receptor: localization on the human X chromosome and evidence for homology with the Tfm locus in the mouse. Proc Natl Acad Sci U S A. 1981 Oct;78(10):6339–6343. doi: 10.1073/pnas.78.10.6339. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Moguilewsky M., Philibert D. RU 38486: potent antiglucocorticoid activity correlated with strong binding to the cytosolic glucocorticoid receptor followed by an impaired activation. J Steroid Biochem. 1984 Jan;20(1):271–276. doi: 10.1016/0022-4731(84)90216-4. [DOI] [PubMed] [Google Scholar]
  23. Munck A., Foley R. Activation of steroid hormone-receptor complexes in intact target cells in physiological conditions. Nature. 1979 Apr 19;278(5706):752–754. doi: 10.1038/278752a0. [DOI] [PubMed] [Google Scholar]
  24. Murakami T., Brandon D., Rodbard D., Loriaux D. L., Lipsett M. B. Glucocorticoid receptor in circulating mononuclear leukocytes. Endocrinology. 1979 Feb;104(2):500–505. doi: 10.1210/endo-104-2-500. [DOI] [PubMed] [Google Scholar]
  25. Olsen K. L. Effects of 17 beta-hydroxy-17 alpha-methyl-estra-4,9,11-triene-3-one (R 1881): evidence for direct involvement of androgens in the defeminization of behaviour in rats. J Endocrinol. 1983 Sep;98(3):431–438. doi: 10.1677/joe.0.0980431. [DOI] [PubMed] [Google Scholar]
  26. Peterson R. E., Imperato-McGinley J., Gautier T., Sturla E. Male pseudohermaphroditism due to steroid 5-alpha-reductase deficiency. Am J Med. 1977 Feb;62(2):170–191. doi: 10.1016/0002-9343(77)90313-8. [DOI] [PubMed] [Google Scholar]
  27. Pinsky L., Kaufman M., Gil-Esteban C., Sumbulian D. Regulation of the androgen receptor in human genital skin fibroblasts, with a review of sex steroid receptor regulation by homologous and heterologous steroids. Can J Biochem Cell Biol. 1983 Jul;61(7):770–778. doi: 10.1139/o83-097. [DOI] [PubMed] [Google Scholar]
  28. Pinsky L., Kaufman M., Summitt R. L. Congenital androgen insensitivity due to a qualitatively abnormal androgen receptor. Am J Med Genet. 1981;10(1):91–99. doi: 10.1002/ajmg.1320100111. [DOI] [PubMed] [Google Scholar]
  29. Pinsky L., Miller J., Shanfield B., Watters G., Wolfe L. S. GM1 gangliosidosis in skin fibroblast culture: enzymatic differences between types 1 and 2 and observations on a third variant. Am J Hum Genet. 1974 Sep;26(5):563–577. [PMC free article] [PubMed] [Google Scholar]
  30. Rochefort H., Borgna J. L. Differences between oestrogen receptor activation by oestrogen and antioestrogen. Nature. 1981 Jul 16;292(5820):257–259. doi: 10.1038/292257a0. [DOI] [PubMed] [Google Scholar]
  31. Seeley D. H., Costas P. D. Transformation of a rabbit progesterone receptor from an 8S form to 5.5S and 4S forms. Mol Cell Endocrinol. 1983 May;30(2):161–178. doi: 10.1016/0303-7207(83)90045-x. [DOI] [PubMed] [Google Scholar]
  32. Weichman B. M., Notides A. C. Estradiol-binding kinetics of the activated and nonactivated estrogen receptor. J Biol Chem. 1977 Dec 25;252(24):8856–8862. [PubMed] [Google Scholar]
  33. Wilson J. D. Dihydrotestosterone formation in cultured human fibroblasts. Comparison of cells from normal subjects and patients with familial incomplete male pseudohermaphroditism, Type 2. J Biol Chem. 1975 May 10;250(9):3498–3504. [PubMed] [Google Scholar]
  34. Wilson J. D., Harrod M. J., Goldstein J. L., Hemsell D. L., MacDonald P. C. Familial incomplete male pseudohermaphroditism, type 1. Evidence for androgen resistance and variable clinical manifestations in a family with the Reifenstein syndrome. N Engl J Med. 1974 May 16;290(20):1097–1103. doi: 10.1056/NEJM197405162902001. [DOI] [PubMed] [Google Scholar]

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