Interaction of human estrogen receptors α and β with the same naturally occurring estrogen response elements

doi:10.1016/S0006-2952(98)00355-4

Biochemical Pharmacology

Volume 57, Issue 6, 15 March 1999, Pages 597-601

https://doi.org/10.1016/S0006-2952(98)00355-4 Get rights and content

Abstract

Estrogen receptors are derived from two different gene products referred to as estrogen receptor-α (ER-α) and ER-β. Both receptors bind to the consensus estrogen response element (ERE) present in the vitellogenin gene, but their binding to hormone response elements present in other estrogen responsive genes has not been reported yet. Using in vitro expressed human receptors, we now show that ER-β binds to a panel of six endogenous hormone response elements (vitellogenin, c-fos, c-jun, pS2, cathepsin D, and choline acetyltransferase) already known to bind ER-α and confer estrogen inducibility to reporter constructs. Binding of ER-α and ER-β occurred at similar DNA concentrations for some EREs, but different DNA concentrations were required to form complexes of the two receptors with other elements. These results illustrate for the first time by direct receptor–DNA binding studies that both ER-α and ER-β bind to a number of EREs present in endogenous hormone regulated genes, and further suggest that the two forms of the receptor display different patterns of affinities for naturally occurring hormone response elements.

Section snippets

Materials

Enzymes were obtained from Boehringer Mannheim, and oligonucleotides were synthesized by Genosys or National Biosciences. Recombinant human ER-α (66 kDa) and ER-β (53 kDa) [2] were obtained from Panvera. Polyacrylamide gel electrophoresis supplies were obtained from Bio-Rad. Antibody H-222 was provided by Dr. Geoffrey Greene of the University of Chicago. All other materials used were of the highest grade commercially available.

Oligonucleotide preparation and gel-shift assays

The nucleotides containing the natural EREs (underlined) used are

Results

As an initial positive control, we tested the ability of the receptors to bind specifically to the vit-ERE as previously reported 8, 16, and to determine the usable range of ER protein concentration. As shown in Fig. 1 , both ER-α and ER-β bound the vit-ERE at protein levels between 25 and 100 ng under our assay conditions. Binding of both receptors was specific as indicated by competition with a 100-fold excess of unlabeled DNA. Note also that the migration of the ER-β complex was slightly

Discussion

We have examined the binding of ER-α and ER-β to a variety of naturally occurring EREs including: (a) the palindromic vitellogenin element, (b) four elements (fos, jun, pS2, and cathepsin D) of the type most frequently observed in endogenous hormone-responsive genes, i.e. one perfect half-site and a second half-site with 1–3 base changes from the palindromic vit-ERE, and (c) a non-palindromic element (choline acetyltransferase) with overall similarity to the vit-ERE (eight of ten nucleotides

Acknowledgements

This work was supported by NIH Grant HD-08615. We would like to thank Ms. Heidi Porter for editorial assistance.

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Biochemical Pharmacology

Abstract

Section snippets

Materials

Oligonucleotide preparation and gel-shift assays

Results

Discussion

Acknowledgements

References (18)

ERβIdentification and characterization of a novel human estrogen receptor

FEBS Lett

Estrogen receptors α and β form heterodimers on DNA

J Biol Chem

Identification of an estrogen response element in the 3′-flanking region of the murine c-fos protooncogene

J Biol Chem

The protooncogene c-jun contains an unusual estrogen-inducible enhancer within the coding sequence

J Biol Chem

In vitro interaction of uterine estrogen receptor with the estrogen response element present in the 3′-flanking region of the murine c-fos protooncogene

J Steroid Biochem Mol Biol

Equilibrium binding of estrogen receptor with DNA using fluorescence anisotropy

J Biol Chem

Cloning of a novel estrogen receptor expressed in rat prostate and ovary

Proc Natl Acad Sci USA

Cloning, chromosomal localization, and functional analysis of the murine estrogen receptor β

Mol Endocrinol

The novel estrogen receptor-β subtypePotential role in the cell- and promoter-specific actions of estrogens and anti-estrogens

FEBS Lett

Cited by (90)

Previous estradiol treatment during midlife maintains transcriptional regulation of memory-related proteins by ERα in the hippocampus in a rat model of menopause

Long-term consequences of estrogens administered in midlife on female cognitive aging

In vitro and in vivo effects of phytoestrogens on protein turnover in rainbow trout (Oncorhynchus mykiss) white muscle

Gene Expression Profiles in Breast Cancer to Identify Estrogen Receptor Target Genes

Study sequence rules of estrogen receptor α-DNA interactions using dual polarization interferometry and computational modeling

Sex Hormone Receptors in Breast Cancer

Rapid CommunicationsInteraction of human estrogen receptors α and β with the same naturally occurring estrogen response elements

Abstract

Section snippets

Materials

Oligonucleotide preparation and gel-shift assays

Results

Discussion

Acknowledgements

FEBS Lett

J Biol Chem

J Biol Chem

J Biol Chem

J Steroid Biochem Mol Biol

J Biol Chem

Cloning of a novel estrogen receptor expressed in rat prostate and ovary

Proc Natl Acad Sci USA

Cloning, chromosomal localization, and functional analysis of the murine estrogen receptor β

Mol Endocrinol

The novel estrogen receptor-β subtypePotential role in the cell- and promoter-specific actions of estrogens and anti-estrogens

FEBS Lett

Rapid Communications
Interaction of human estrogen receptors α and β with the same naturally occurring estrogen response elements