DNA binding by estrogen receptor-alpha is essential for the transcriptional response to estrogen in the liver and the uterus

Mol Endocrinol. 2009 Oct;23(10):1544-55. doi: 10.1210/me.2009-0045. Epub 2009 Jul 2.

Abstract

The majority of the biological effects of estrogens in the reproductive tract are mediated by estrogen receptor (ER)alpha, which regulates transcription by several mechanisms. Because the tissue-specific effects of some ERalpha ligands may be caused by tissue-specific transcriptional mechanisms of ERalpha, we aimed to identify the contribution of DNA recognition to these mechanisms in two clinically important target organs, namely uterus and liver. We used a genetic mouse model that dissects DNA binding-dependent vs. independent transcriptional regulation elicited by ERalpha. The EAAE mutant harbors amino acid exchanges at four positions of the DNA-binding domain (DBD) of ERalpha. This construct was knocked in the ERalpha gene locus to produce ERalpha((EAAE/EAAE)) mice devoid of a functional ERalpha DBD. The phenotype of the ERalpha((EAAE/EAAE)) mice resembles the general loss-of-function phenotype of alphaER knockout mutant mice with hypoplastic uteri, hemorrhagic ovaries, and impaired mammary gland development. In agreement with this phenotype, the expression pattern of the ERalpha((EAAE/EAAE)) mutant mice in liver obtained by genome-wide gene expression profiling supports the observation of a near-complete loss of estrogen-dependent gene regulation in comparison with the wild type. Further gene expression analyses to validate the results of the microarray data were performed by quantitative RT-PCR. The analyses indicate that both gene activation and repression by estrogen-bound ERalpha rely on an intact DBD in vivo.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Autophagy-Related Proteins
  • DNA / metabolism*
  • Estrogen Receptor alpha / chemistry
  • Estrogen Receptor alpha / metabolism*
  • Estrogens / pharmacology*
  • Ethinyl Estradiol / pharmacology
  • Female
  • Gene Expression Regulation / drug effects
  • HeLa Cells
  • Humans
  • Infertility, Female / genetics
  • Interleukin-1beta / pharmacology
  • Liver / drug effects
  • Liver / metabolism*
  • Mice
  • Mice, Mutant Strains
  • Molecular Sequence Data
  • Mutation / genetics
  • Protein Binding / drug effects
  • Protein Structure, Tertiary
  • Protein-Serine-Threonine Kinases / metabolism
  • Proteins / metabolism
  • Repressor Proteins / metabolism
  • Response Elements / genetics
  • Transcription, Genetic / drug effects*
  • Uterus / drug effects
  • Uterus / metabolism*

Substances

  • Autophagy-Related Proteins
  • Estrogen Receptor alpha
  • Estrogens
  • Interleukin-1beta
  • Lix1 protein, mouse
  • Proteins
  • Repressor Proteins
  • Ethinyl Estradiol
  • DNA
  • Plk3 protein, mouse
  • Protein-Serine-Threonine Kinases