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J Biol Chem, Vol. 274, Issue 45, 32099-32107, November 5, 1999
From the bcl-2 gene expression is induced by
17
Mechanisms of Transcriptional Activation of bcl-2
Gene Expression by 17
-Estradiol in Breast Cancer Cells
,,,,
,,, and
Department of Veterinary Physiology and
Pharmacology and Department of Biochemistry and Biophysics,
Texas A&M University, College Station, Texas 77843 and the
§ Burnham Institute, La Jolla, California 92037
-estradiol (E2) in T47D and MCF-7 human breast cancer cells, and
the mechanism of E2 responsiveness was further investigated by analysis
of the bcl-2 gene promoter. The 
1602 to 1534 distal
region (bcl-2j) of the promoter was E2-responsive; however, in gel
mobility shift assays, the estrogen receptor 
(ER
)
did not bind [32P]bcl-2j, whereas Sp1 protein formed a
retarded band complex. Further analysis demonstrated that the upstream
region (1603 to 1579) of the bcl-2 gene promoter
contained two GC/GA-rich sites at 1601 (5'-GGGCTGG-3') and 1588
(3'-GGAGGG-5') that bound Sp1 protein. Subsequent studies confirmed
that transactivation by E2 was dependent on ER/Sp1
interactions with both GC-rich sites, and this was confirmed by
in vitro footprinting. In contrast, a 21-base pair
E2-responsive downstream region (1578 to 1534) did not bind Sp1 or
ER protein; however, analysis of a complex binding
pattern with nuclear extracts showed that ATF-1 and CREB-1 bound to
this motif. These data coupled with results of transient transfection
studies demonstrated that transcriptional activation by E2 of the
1578 to 1534 region of the bcl-2 gene promoter was
dependent on induction of cAMP and subsequent activation through a cAMP
response element. Thus, hormone regulation of bcl-2 gene expression in breast cancer cells involves multiple enhancer elements and E2-mediated transactivation does not require direct binding of the
estrogen receptor with promoter DNA.
Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.
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