title: DNA binding by estrogen receptor-alpha is essential for the transcriptional response to estrogen in the liver and the uterus.
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.
authors: Bayer Schering Pharma AG, Therapeutic Research Group Women's Healthcare, Müllerstrasse 178, 13353 Berlin, Germany.
authors: Dörthe L
authors: DL
authors: Ahlbory-Dieker
authors: Brenda D
authors: BD
authors: Stride
authors: Gabriele
authors: G
authors: Leder
authors: Jenny
authors: J
authors: Schkoldow
authors: Susanne
authors: S
authors: Trölenberg
authors: Henrik
authors: H
authors: Seidel
authors: Christiane
authors: C
authors: Otto
authors: Anette
authors: A
authors: Sommer
authors: Malcolm G
authors: MG
authors: Parker
authors: Günther
authors: G
authors: Schütz
authors: Tim M
authors: TM
authors: Wintermantel
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000071183:Autophagy-Related Proteins; D004247:DNA / Q000378:metabolism*; D047628:Estrogen Receptor alpha / Q000737:chemistry / Q000378:metabolism*; D004967:Estrogens / Q000494:pharmacology*; D004997:Ethinyl Estradiol / Q000494:pharmacology; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects; D006367:HeLa Cells; D006801:Humans; D007247:Infertility, Female / Q000235:genetics; D053583:Interleukin-1beta / Q000494:pharmacology; D008099:Liver / Q000187:drug effects / Q000378:metabolism*; D051379:Mice; D008817:Mice, Mutant Strains; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D011485:Protein Binding / Q000187:drug effects; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000378:metabolism; D012097:Repressor Proteins / Q000378:metabolism; D020218:Response Elements / Q000235:genetics; D014158:Transcription, Genetic / Q000187:drug effects*; D014599:Uterus / Q000187:drug effects / Q000378:metabolism*
chemical_list: D000071183:Autophagy-Related Proteins; D047628:Estrogen Receptor alpha; D004967:Estrogens; D053583:Interleukin-1beta; C499180:Lix1 protein, mouse; D011506:Proteins; D012097:Repressor Proteins; D004997:Ethinyl Estradiol; D004247:DNA; C093328:Plk3 protein, mouse; D017346:Protein Serine-Threonine Kinases