Estrogen-responsive element of the human pS2 gene is an imperfectly palindromic sequence.

Abstract

Using chimeric recombinants transfected into HeLa cells and a transient expression assay, we demonstrate that the 5'-flanking region of the pS2 gene from position -428 to position -324 exhibits both constitutive and estrogeninducible enhancer activity. The estrogen-inducible activity, but not the constitutive activity, was inhibited by antiestrogens.ICI 164,384 behaved as a pure antagonist, whereas hydroxytamoxifen was a partial agonist-antagonist. The estrogenresponsive element of the pS2 gene has been narrowed down by site-directed deletion mutagenesis to a 13-base-pair (position -405 to position -393) imperfectly palindromic sequence, which in isolation can confer estrogen inducibility to the heterologous rabbit IB-globin gene promoter. On the other hand, the sequences responsible for the constitutive enhancer activity are spread over the entire region.The human pS2 gene was initially characterized as a gene whose expression is specifically controlled by estrogen in the breast cancer cell line MCF-7 (1, 2). The increase in pS2 mRNA after addition of estradiol to the culture medium is a primary transcriptional event (3), suggesting that control of pS2 gene promoter activity by the estrogen receptor is mediated by a cis-acting estrogen-responsive element (ERE) that could be located in the 5'-flanking region ofthe pS2 gene. Using MCF-7 cells transformed with chimeric recombinants containing the promoter region of the pS2 gene controlling transcription of the gene conferring resistance to neomycin (G418), it has indeed been shown that the 5'-flanking region of the pS2 gene from position approximately -3000 to position +10 contains an estrogen-responsive promoter (4). Transfection of these recombinants together with the human estrogen receptor expression vector HEO in the presence or absence of estradiol led to the hypothesis that both constitutive and estrogen-inducible enhancer elements are located within the region of the pS2 gene from position -428 to position -332 (7).In the present study we demonstrate that the pS2 gene sequence from position -428 to position -324 indeed contains both constitutive and estrogen-inducible enhancer elements. Furthermore, the exact location of the ERE has been determined by using both deletion and point mutagenesis. We show that the pS2 ERE sequence is related to those of characterized EREs but that it is not a perfect palindrome, which results in a weaker ERE. Fig. 3A (note the presence Abbreviation: ERE, estrogen-responsive element. MATERIALS AND METHODS 1218The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.

Effects of the Environmental Oestrogens Bisphenol A, Tetrachlorobisphenol A, Tetrabromobisphenol A, 4‐Hydroxybiphenyl and 4,4′‐Dihydroxybiphenyl on Oestrogen Receptor Binding, Cell Proliferation and Regulation of Oestrogen Sensitive Proteins in the Human Breast Cancer Cell Line MCF‐7

“…However, in contrast to pS2 and PgR, changes in cytosolic oestrogen receptors is most probably regulated by both transcriptional and post-transcriptional mechanisms. The promoter of the pS2 (Berry et al 1989) and PgR gene (Kraus et al 1994) contains an oestrogen receptor response element, although somewhat imperfect. The promoter of the oestrogen receptor gene, however, lacks the canonical oestrogen ERE (Piva et al 1992).…”

Identification of estrogen-responsive genes based on the DNA binding properties of estrogen receptors using high-throughput sequencing technology

“…However, among them, only a small number have been shown to possess functional EREs within the transcription regulatory region. In mammals, these genes include transcription factors, such as JUN [32] , FOS [33] , PGR [34] , and TP53 [35] , intracellular signaling molecules, such as HRAS [36] , BCL2 [37] , and BRCA1 [38] , enzymes, such as CHAT [39] , NQO1 [40] , and CKB [41] , secreted proteins, such as LTF [42] , SCGB1A1 [43] , OVGP1 [44] , C3 [45] , and AGT [46] , hormones, such as LHB [47] , OXT [48] , PRL [49] , and AVP [50] , membrane proteins, such as SNAT2 [51] and VEGFA [52] , the motogen TFF1 [53] , and the protease CTSD [54] . These genes are assumed to directly mediate various estrogen actions in normal tissues, as well as in cancer and other diseases.…”

Recruitment of Distinct Chromatin-modifying Complexes by Tamoxifen-complexed Estrogen Receptor at Natural Target Gene Promoters in Vivo

“…To explore the mechanisms underlying the inhibitory effects of OHT, we employed human MCF-7 breast cancer cells, which are enriched in ER␣. We initially analyzed the effects of OHT on the expression of two previously characterized E-regulated genes, pS2 and c-Myc (19,20). In the experiment described in Fig.…”