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Human Estrogen Receptor (ER) Gene Promoter-P1: Estradiol-Independent Activity and Estradiol Inducibility in ER⁺ and ER^- Cells

Laboratoire de Virologie Moléculaire (I.T., N.P., A.S.) Unité INSERM U412 Ecole Normale Supérieure de Lyon 69364 Lyon Cédex 07, France
Département d’Anatomie et de Cytologie Pathologiques (I.T., N.P.) Centre Léon Bérard 69373 Lyon Cédex 08, France
Division of Neoplastic Disease Mechanisms (M.B.) Dana Farber Cancer Institute Boston, MA 02115

Estrogen receptor (ER) is expressed at a low level in normal tissues such as breast and uterus but at a high level in breast and endometrial carcinomas. A proximal element (ERF-1) located between positions +133 and +204 relative to the promoter P1 major initiation site has been recently identified in ER⁺ cells and contributes to the differential promoter activity between ER⁺ and ER^- cells. In this study, MCF7 and HeLa cells were transfected with chloramphenicol acetyltransferase constructs containing ER gene promoter P1 sequences. We show here that the sequences lying between nucleotides +13 to +212 are also essential for transcription at the ER gene promoter P1 in ER^- cells, which do not express ERF-1. Interestingly, on gel shift experiments, a complex specific to ER^- cells forms in the region spanning nucleotides +123 to +210. We also show that promoter P1 is responsive to estradiol in cells expressing endogenous (MCF7) or exogenous ER. We further demonstrate, using mutational analysis and gel retardation assays, that the three half-estrogen response elements located between nucleotides -420 and -892 are responsible for the estradiol inducibility of promoter P1. Because estradiol has a mitogenic effect on both breast and endometrial epithelial cells, our data would give an insight into the role of estrogens in the occurence of breast and endometrial carcinomas.

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