Genomic binding-site cloning reveals an estrogen-responsive gene that encodes a RING finger protein

Department of Biochemistry, Saitama Medical School, Japan.
Proceedings of the National Academy of Sciences (Impact Factor: 9.67). 01/1994; 90(23):11117-21. DOI: 10.1073/pnas.90.23.11117
Source: PubMed


Estrogen receptor (ER)-binding fragments were isolated from human genomic DNA by using a recombinant ER protein. Using one of these fragments as a probe, we have identified an estrogen-responsive gene that encodes a putative zinc finger protein. It has a RING finger motif present in a family of apparent DNA-binding proteins and is designated estrogen-responsive finger protein (efp). efp cDNA contains a consensus estrogen-responsive element at the 3' untranslated region that can act as a downstream estrogen-dependent enhancer. Moreover, efp is regulated by estrogen as demonstrated at both the mRNA and the protein level in ER-positive cells derived from mammary gland. These data suggest that efp may represent an estrogen-responsive transcription factor that mediates phenotypic expression of the diverse estrogen action. Thus, the genomic binding-site cloning may be applicable for isolation of the target genes of other transcription factors.

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Available from: Akira Orimo
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    • "Identification of the target genes of transcription factors is critical to our understanding of transcriptional networks. For example, as canonical EREs were rarely found in promoter regions (O'Lone et al. 2004), it was difficult to identify the target genes of ERs by sequence motifs alone; filter binding (Inoue et al. 1993), computational approaches (Bourdeau et al. 2004; Kamalakaran et al. 2005) and ChIP on Chip (Carroll et al. 2005; Laganiere et al. 2005) have been applied to their identification . As these studies focused primarily on cultured cells, no information regarding whole organisms was available. "
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    ABSTRACT: Estrogens play a central role in the reproduction of vertebrates and affect a variety of biological processes. The major target molecules of estrogens are nuclear estrogen receptors (ERs), which have been studied extensively at the molecular level. In contrast, our knowledge of the genes that are regulated directly by ERs remains limited, especially at the level of the whole organism rather than cultured cells. In order to identify genes that are regulated directly by ERs in vivo, we used estrogen treated mouse uterus and performed chromatin immunoprecipitation. Sequence analysis of a precipitated DNA fragment enabled alignment with the mouse genomic sequence and revealed that the promoter region of the gene encoding aquaporin 5 (AQP5) was precipitated with antibody against ER alpha. Quantitative PCR and DNA microarray analyses confirmed that AQP5 is activated soon after administration of estrogen. In addition, the promoter region of AQP5 contained a functional estrogen response element that was activated directly by estrogen. Although several AQP genes are expressed in the uterus, only direct activation of AQP5 could be detected following treatment with estrogen. This chromatin immunoprecipitation-mediated target identification may be applicable to the study of other transcription factor networks.
    Full-text · Article · Nov 2006 · Genes to Cells
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    • ".edu/ERTargetDB/SupplementaryInfo.pdf). For example, the reported ERE motifs GGTCATGGT- GACC in the human estrogen-responsive gene (efp) that encodes a RING finger protein (Inoue et al. 1993) and GGACACCATCTGTCC in the rat luteinizing hormone gene (Lhb) (Shupnik et al. 1989) were not found in the corresponding 7 kb promoter region. The 32 ER target genes that belong to the ER/X pathway of human, mouse and rat are reported to use the 'tethering,' mechanism with AP-1, Sp1 or NF-by individual laboratories (Supplementary Table 2 available at "
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    ABSTRACT: The estrogen receptor (ER) plays an important role in several physiologic functions of both the reproductive and non-reproductive systems. Malignancies of the ER have been associated with the development of cancers, including those of the prostate and breast. Hence it has become of significant importance to characterize the transcriptional regulation of ER target genes. We have created ERTargetDB in order to integrate the previously published ER target gene information that is available in various publications and databases. This information resource provides researchers with an easy access to ER target genes and the regulatory mechanisms in the corresponding promoters. The current version contains 40 genes with experimentally verified estrogen response elements (EREs), 32 experimentally verified ERE tethering sites, 40 genes identified by the chromatin immunoprecipitation microarray, 381 genes from gene expression microarray and 2948 genes from computational prediction. ERTargetDB provides an integral information resource for direct target genes of ERs for the endocrinology research community. It should prove useful in the investigation of gene regulation and aid the development of computational tools for the prediction of ER target genes.
    Full-text · Article · Nov 2005 · Journal of Molecular Endocrinology
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    • "The consensus ERE is a 13-bp palindromic sequence containing two inverted repeats of 5′-GGATC-3′ separated by three base pairs. Although perfect EREs have only been discovered in two human estrogen-regulated genes (cytochrome c oxidase subunit VIIa-related protein (COX7RP) (Watanabe et al. 1998) and estrogen responsive finger protein (Efp) (Inoue et al. 1993)), ERα can bind to non-perfect or half ERE sequences, particularly in the context of appropriate flanking sequences. ERα can also affect "
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    ABSTRACT: Estrogen receptor alpha (ERalpha) has an established role in promoting breast cancer. Transcriptional activation by ERalpha is a complex and multistep process, and it is influenced by coactivator and corepressor proteins that can either positively or negatively modulate ERalpha-mediated transcriptional activity. Corepressors are proposed to provide a counterbalance to the estrogen-induced transactivation, and represent a potential mechanism employed by the cell to regulate hormonal responses. In this review, we present evidence from tissue culture, animal and clinical studies, supporting the hypothesis that corepressors are crucial regulators of ERalpha-mediated action, and that their loss could promote breast cancer development and resistance to endocrine therapy. We propose that ERalpha corepressors play an important biological role by controlling the magnitude of the estrogen response, mediating antiestrogen inhibition of ERalpha, repressing DNA-bound ERalpha in the absence of the ligand, and conferring active repression of ERalpha-downregulated genes. Different ERalpha corepressors regulate steroid receptor activity through a variety of mechanisms, including formation of multiprotein complexes that are able to affect chromatin remodeling, histone deacetylation, or basal transcription. Other mechanisms include competition with coactivators, interference with DNA binding and ERalpha homodimerization, alteration of ERalpha stability, sequestration of ERalpha in the cytoplasm, and effects on RNA processing. Most ERalpha corepressors can control the receptor's activity through more than one mechanism, and it is possible that the synergy between different pathways cooperates to fully inhibit ERalpha transcriptional activity, and create an integrated response to a variety of different cellular signaling pathways. We will discuss the role of corepressors in tumor suppression and the link they might present between ERalpha regulation and DNA repair. Finally, we will discuss major challenges in the field and speculate on the exciting findings that await us in the next few years.
    Preview · Article · Jan 2004 · Endocrine Related Cancer
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