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Volume 272, Number 36, Issue of September 5, 1997 pp. 22425-22431
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.

Cloning and Characterization of the Type I Inositol 1,4,5-Trisphosphate Receptor Gene Promoter
REGULATION BY 17-ESTRADIOL IN OSTEOBLASTS

(Received for publication, April 10, 1997)

Keith L. Kirkwood ^§ , Kristen Homick ^¶ , Marc B. Dragon ^¶ and Peter G. Bradford ^§

From the  Department of Oral Biology, School of Dental Medicine, ^§ Center for the Molecular Mechanisms of Disease and Aging, ^¶ Howard Hughes Medical Institute Undergraduate Biological Sciences Education Program, and  Department of Pharmacology and Toxicology, School of Medicine and Biomedical Sciences, State University of New York, Buffalo, New York 14214-3000

The inositol 1,4,5-trisphosphate (InsP₃) receptor is essential for signal Ca²⁺ release from intracellular stores and for capacitative Ca²⁺ entry. We have isolated the promoter and proximal DNA segments of the human type I InsP₃ receptor gene. Transcription initiation in human G-292 osteosarcoma and HL-60 promyelocytic leukemia cells was shown to occur predominantly from an adenine residue located 39 base pairs downstream of a consensus TATA box element. Upstream DNA including the TATA box promoted directional transcription of a chloramphenicol acetyltransferase reporter gene when transfected into G-292 cells. A negative regulatory element in the distal promoter and a positive element in the proximal region were identified by deletion mapping and transcription assays. The proximal region enhanced transcription in response to 12-O-tetradecanoylphorbol-13-acetate or serum, but conferred transcriptional repression in response to 1,25-dihydroxyvitamin D₃ or 17-estradiol. The repressive effect of 17-estradiol was mediated by the nuclear estrogen receptor, as estrogen-dependent transcriptional repression was inhibited by the antiestrogen tamoxifen and the estrogen receptor antagonist ICI 182,780. This is the first study of the type I InsP₃ receptor gene promoter, and the results suggest a mechanism by which chronic estrogen treatment of osteoblasts affects type I InsP₃ receptor gene expression, signal transduction, and secretion.

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