Amino acid substitutions in the hormone-binding domain of the human androgen receptor alter the stability of the hormone receptor complex

Marco Marcelli, Sonia Zoppi, Carol M. Wilson, Jim Griffin III, Michael J. McPhaul

Research output: Contribution to journalArticle

47 Citations (Scopus)

Abstract

We have investigated the basis of androgen resistance in seven unrelated individuals with complete testicular feminization or Reifenstein syndrome caused by single amino acid substitutions in the hormone-binding domain of the androgen receptor. Monolayer-binding assays of cultured genital skin fibroblasts demonstrated absent ligand binding, qualitative abnormalities of androgen binding, or a decreased amount of qualitatively normal receptor. The consequences of these mutations were examined by introducing the mutations by site-directed mutagenesis into the androgen receptor cDNA sequence and expressing the mutant cDNAs in mammalian cells. The effects of the amino acid substitutions on the binding of different androgens and on the capacity of the ligand-bound receptors to activate a reporter gene were investigated. Substantial differences were found in the responses of the mutant androgen receptors to incubation with testosterone, 5α-dihydrotestosterone, and mibolerone. In several instances, increased doses of hormone or increased frequency of hormone addition to the incubation medium resulted in normal or near normal activation of a reporter gene by cells expressing the mutant androgen receptors. These studies suggest that the stability of the hormone receptor complex is a major determinant of receptor function in vivo.

Original languageEnglish (US)
Pages (from-to)1642-1650
Number of pages9
JournalJournal of Clinical Investigation
Volume94
Issue number4
DOIs
StatePublished - Jan 1 1994

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Androgen Receptors
Amino Acid Substitution
Hormones
Androgen-Insensitivity Syndrome
Androgens
Reporter Genes
Complementary DNA
Ligands
Mutation
Dihydrotestosterone
Site-Directed Mutagenesis
Testosterone
Fibroblasts
Skin
human AR protein

Keywords

  • androgen
  • hormone resistance
  • insensitivity
  • receptor

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Amino acid substitutions in the hormone-binding domain of the human androgen receptor alter the stability of the hormone receptor complex. / Marcelli, Marco; Zoppi, Sonia; Wilson, Carol M.; Griffin III, Jim; McPhaul, Michael J.

In: Journal of Clinical Investigation, Vol. 94, No. 4, 01.01.1994, p. 1642-1650.

Research output: Contribution to journalArticle

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AB - We have investigated the basis of androgen resistance in seven unrelated individuals with complete testicular feminization or Reifenstein syndrome caused by single amino acid substitutions in the hormone-binding domain of the androgen receptor. Monolayer-binding assays of cultured genital skin fibroblasts demonstrated absent ligand binding, qualitative abnormalities of androgen binding, or a decreased amount of qualitatively normal receptor. The consequences of these mutations were examined by introducing the mutations by site-directed mutagenesis into the androgen receptor cDNA sequence and expressing the mutant cDNAs in mammalian cells. The effects of the amino acid substitutions on the binding of different androgens and on the capacity of the ligand-bound receptors to activate a reporter gene were investigated. Substantial differences were found in the responses of the mutant androgen receptors to incubation with testosterone, 5α-dihydrotestosterone, and mibolerone. In several instances, increased doses of hormone or increased frequency of hormone addition to the incubation medium resulted in normal or near normal activation of a reporter gene by cells expressing the mutant androgen receptors. These studies suggest that the stability of the hormone receptor complex is a major determinant of receptor function in vivo.

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