Testosterone and 5α-dihydrotestosterone interact differently with the androgen receptor to enhance transcription of the MMTV-CAT reporter gene

J. P. Deslypere, M. Young, J. D. Wilson, M. J. McPhaul

Research output: Contribution to journalArticle

166 Citations (Scopus)

Abstract

Testosterone and its 5α-reduced derivative 5α-dihydrotestosterone exert different actions in the male during embryogenesis and in postnatal life. Nevertheless the two hormones bind to the same intracellular androgen receptor, and genetic and endocrinological studies in the Tfm mouse suggest that the actions of both hormones are mediated by this single receptor. Previous studies indicate that dihydrotestosterone binds more tightly to the androgen receptor but that the Bmax of binding of the two hormones is the same. To determine whether these differences in binding parameters could explain the mechanism by which the two hormones exert different physiological actions via the same receptor, we introduced a plasmid encoding the androgen receptor cDNA and a reporter plasmid encoding MMTV-CAT into Chinese hamster ovary cells. These cells do not express endogenous androgen receptor and do not convert testosterone to dihydrotestosterone. Therefore, it was possible to examine the relation between the concentration of each of the steroids and reporter gene expression. Both hormones enhanced CAT activity, but dihydrotestosterone was approximately 10 times as potent (half maximal of 0.018 nM) as testosterone (half maximal of 0.2 nM); the maximal activity achieved was the same for the two androgens. These findings are nearly identical to the apparent Kd values for the interaction of the two hormones with the androgen receptor. Although testosterone and dihydrotestosterone may influence the expression of other genes differently, these findings are compatible with a model system in which the differential effects can be explained as a consequence of different binding affinities to the receptor. In peripheral target tissues dihydrotestosterone formation would be necessary for androgen action whereas in tissues in which testosterone concentration is high (testes, wolffian ducts) dihydrotestosterone formation may not be essential for virilization.

Original languageEnglish (US)
Pages (from-to)15-22
Number of pages8
JournalMolecular and Cellular Endocrinology
Volume88
Issue number1-3
DOIs
StatePublished - 1992

Fingerprint

Dihydrotestosterone
Androgen Receptors
Transcription
Reporter Genes
Testosterone
Genes
Hormones
Androgens
Plasmids
Wolffian Ducts
Tissue
Virilism
Gene Expression
Cricetulus
Gene expression
Ducts
Embryonic Development
Testis
Ovary
Complementary DNA

Keywords

  • 5α-Dihydrotestosterone
  • Androgen receptor
  • MMTV-CAT reporter gene
  • Testosterone
  • Transcription

ASJC Scopus subject areas

  • Endocrinology
  • Endocrinology, Diabetes and Metabolism

Cite this

Testosterone and 5α-dihydrotestosterone interact differently with the androgen receptor to enhance transcription of the MMTV-CAT reporter gene. / Deslypere, J. P.; Young, M.; Wilson, J. D.; McPhaul, M. J.

In: Molecular and Cellular Endocrinology, Vol. 88, No. 1-3, 1992, p. 15-22.

Research output: Contribution to journalArticle

@article{7ca0f70fd464453c849abda872f00727,
title = "Testosterone and 5α-dihydrotestosterone interact differently with the androgen receptor to enhance transcription of the MMTV-CAT reporter gene",
abstract = "Testosterone and its 5α-reduced derivative 5α-dihydrotestosterone exert different actions in the male during embryogenesis and in postnatal life. Nevertheless the two hormones bind to the same intracellular androgen receptor, and genetic and endocrinological studies in the Tfm mouse suggest that the actions of both hormones are mediated by this single receptor. Previous studies indicate that dihydrotestosterone binds more tightly to the androgen receptor but that the Bmax of binding of the two hormones is the same. To determine whether these differences in binding parameters could explain the mechanism by which the two hormones exert different physiological actions via the same receptor, we introduced a plasmid encoding the androgen receptor cDNA and a reporter plasmid encoding MMTV-CAT into Chinese hamster ovary cells. These cells do not express endogenous androgen receptor and do not convert testosterone to dihydrotestosterone. Therefore, it was possible to examine the relation between the concentration of each of the steroids and reporter gene expression. Both hormones enhanced CAT activity, but dihydrotestosterone was approximately 10 times as potent (half maximal of 0.018 nM) as testosterone (half maximal of 0.2 nM); the maximal activity achieved was the same for the two androgens. These findings are nearly identical to the apparent Kd values for the interaction of the two hormones with the androgen receptor. Although testosterone and dihydrotestosterone may influence the expression of other genes differently, these findings are compatible with a model system in which the differential effects can be explained as a consequence of different binding affinities to the receptor. In peripheral target tissues dihydrotestosterone formation would be necessary for androgen action whereas in tissues in which testosterone concentration is high (testes, wolffian ducts) dihydrotestosterone formation may not be essential for virilization.",
keywords = "5α-Dihydrotestosterone, Androgen receptor, MMTV-CAT reporter gene, Testosterone, Transcription",
author = "Deslypere, {J. P.} and M. Young and Wilson, {J. D.} and McPhaul, {M. J.}",
year = "1992",
doi = "10.1016/0303-7207(92)90004-P",
language = "English (US)",
volume = "88",
pages = "15--22",
journal = "Molecular and Cellular Endocrinology",
issn = "0303-7207",
publisher = "Elsevier Ireland Ltd",
number = "1-3",

}

TY - JOUR

T1 - Testosterone and 5α-dihydrotestosterone interact differently with the androgen receptor to enhance transcription of the MMTV-CAT reporter gene

AU - Deslypere, J. P.

AU - Young, M.

AU - Wilson, J. D.

AU - McPhaul, M. J.

PY - 1992

Y1 - 1992

N2 - Testosterone and its 5α-reduced derivative 5α-dihydrotestosterone exert different actions in the male during embryogenesis and in postnatal life. Nevertheless the two hormones bind to the same intracellular androgen receptor, and genetic and endocrinological studies in the Tfm mouse suggest that the actions of both hormones are mediated by this single receptor. Previous studies indicate that dihydrotestosterone binds more tightly to the androgen receptor but that the Bmax of binding of the two hormones is the same. To determine whether these differences in binding parameters could explain the mechanism by which the two hormones exert different physiological actions via the same receptor, we introduced a plasmid encoding the androgen receptor cDNA and a reporter plasmid encoding MMTV-CAT into Chinese hamster ovary cells. These cells do not express endogenous androgen receptor and do not convert testosterone to dihydrotestosterone. Therefore, it was possible to examine the relation between the concentration of each of the steroids and reporter gene expression. Both hormones enhanced CAT activity, but dihydrotestosterone was approximately 10 times as potent (half maximal of 0.018 nM) as testosterone (half maximal of 0.2 nM); the maximal activity achieved was the same for the two androgens. These findings are nearly identical to the apparent Kd values for the interaction of the two hormones with the androgen receptor. Although testosterone and dihydrotestosterone may influence the expression of other genes differently, these findings are compatible with a model system in which the differential effects can be explained as a consequence of different binding affinities to the receptor. In peripheral target tissues dihydrotestosterone formation would be necessary for androgen action whereas in tissues in which testosterone concentration is high (testes, wolffian ducts) dihydrotestosterone formation may not be essential for virilization.

AB - Testosterone and its 5α-reduced derivative 5α-dihydrotestosterone exert different actions in the male during embryogenesis and in postnatal life. Nevertheless the two hormones bind to the same intracellular androgen receptor, and genetic and endocrinological studies in the Tfm mouse suggest that the actions of both hormones are mediated by this single receptor. Previous studies indicate that dihydrotestosterone binds more tightly to the androgen receptor but that the Bmax of binding of the two hormones is the same. To determine whether these differences in binding parameters could explain the mechanism by which the two hormones exert different physiological actions via the same receptor, we introduced a plasmid encoding the androgen receptor cDNA and a reporter plasmid encoding MMTV-CAT into Chinese hamster ovary cells. These cells do not express endogenous androgen receptor and do not convert testosterone to dihydrotestosterone. Therefore, it was possible to examine the relation between the concentration of each of the steroids and reporter gene expression. Both hormones enhanced CAT activity, but dihydrotestosterone was approximately 10 times as potent (half maximal of 0.018 nM) as testosterone (half maximal of 0.2 nM); the maximal activity achieved was the same for the two androgens. These findings are nearly identical to the apparent Kd values for the interaction of the two hormones with the androgen receptor. Although testosterone and dihydrotestosterone may influence the expression of other genes differently, these findings are compatible with a model system in which the differential effects can be explained as a consequence of different binding affinities to the receptor. In peripheral target tissues dihydrotestosterone formation would be necessary for androgen action whereas in tissues in which testosterone concentration is high (testes, wolffian ducts) dihydrotestosterone formation may not be essential for virilization.

KW - 5α-Dihydrotestosterone

KW - Androgen receptor

KW - MMTV-CAT reporter gene

KW - Testosterone

KW - Transcription

UR - http://www.scopus.com/inward/record.url?scp=0026782628&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0026782628&partnerID=8YFLogxK

U2 - 10.1016/0303-7207(92)90004-P

DO - 10.1016/0303-7207(92)90004-P

M3 - Article

VL - 88

SP - 15

EP - 22

JO - Molecular and Cellular Endocrinology

JF - Molecular and Cellular Endocrinology

SN - 0303-7207

IS - 1-3

ER -