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J. Biol. Chem., Vol. 276, Issue 18, 15519-15526, May 4, 2001
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From the Department of Surgery, Stanford University School of
Medicine, Stanford, California 94305-3229
Expression of human estrogen receptor-
Genomic Structure of the Promoters of the Human Estrogen
Receptor-
Gene Demonstrate Changes in Chromatin Structure Induced by
AP2
*
(ER)
involves the activity from several promoters that give rise to
alternate untranslated 5' exons. However, the genomic locations of the
alternate 5' exons have not been reported previously. We have developed
a contig map of the human ER gene that includes all of the known
alternate 5' exons. By using S1 nuclease and 5'- rapid amplification of cDNA ends, the cap sites for the alternate ER transcripts E and H were identified. DNase I-hypersensitive sites specific to
ER-positive cells were associated with each of the cap sites. A
DNase I-hypersensitive site, HS1, was localized to binding sites for
AP2 in the untranslated region of exon 1 and was invariably present in
the chromatin structure of ER-positive cells. Overexpression of
AP2
in human mammary epithelial cells generated the
HS1-hypersensitive site. The ER promoter was induced by AP2 in
mammary epithelial cells, and trans-activation was dependent upon the
region of the promoter containing the HS1 site. These results
demonstrate that AP2 trans-activates the ER gene in
hormone-responsive tumors by inducing changes in the chromatin
structure of the ER promoter. These data are further evidence for a
critical role for AP2 in the oncogenesis of hormone-responsive breast cancers.
*
This work was supported in part by National Institutes of
Health Grant R01 CA77350.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Supported in part by the George H. A. Clowes, Jr., Memorial
Research Career Development award through the American College of
Surgeons. To whom correspondence should be addressed: MSLS, Rm. P214,
1201 Welch Rd., Stanford University School of Medicine, Stanford, CA
94305-5494. Tel.: 650-723-9799; Fax: 650-724-3229; E-mail:
ronald.weigel@stanford.edu.
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