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Molecular Endocrinology Vol. 3, No. 2 400-408
doi:10.1210/mend-3-2-400
Copyright © 1989 by the Endocrine Society.
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Regulation of Epidermal Growth Factor Receptor Gene Expression

Laurie G. Hudson*, June B. Santon* and Gordon N. Gill

Department of Medicine, Division of Endocrinology and Metabolism, Center for Molecular Genetics, University of California-San Diego School of Medicine La Jolla, California 92093

Address requests for reprints to: Gordon N. Gill, M.D., Department of Medicine, M-013E, University of California-San Diego, La Jolla, California 92093.

Abstract

Synthesis and metabolism of the epidermal growth factor (EGF) receptor are extensively regulated to modulate cellular responses to ligand. To study regulation of EGF receptor gene expression, the 5' region of the gene was isolated from a human placental genomic library. A 5' proximal 1.1-kilobase fragment (–1100 to –19 relative to the ATG translation start site) and subfragments of this were subcloned in both forward and reverse orientations into the luciferase expression vector pSVOAL{Delta}5' and transfected into human cell lines. Luciferase activity was stimulated by treatment of transfected HeLa cells with EGF, 12-O-tetradecanoylphorbol 13-acetate (TPA), (Bu)2 cAMP, retinoic acid, and dexamethasone. Deletion analysis indicated full retention of activity after removal of the –1100 to –485 region (–485 to –19 fragment), but a 5-fold reduction in activity on removal of the –485 to –153 region (–153 to –19 fragment). Despite a reduction in basal activity, the proximal 134-basepair fragment retained responses to all inducers. Additivity was observed in response to maximal concentrations of TPA plus retinoic acid and of TPA plus (Bu)2 cAMP; the response to a combination of four inducers exceeded that to the RSV-LTR strong promoter. Differences in stimulated responses were observed in various recipients, with hepatoma HepG2 cells lacking responses to (Bu)2 cAMP and glioblastoma T98G cells lacking responses to EGF and TPA. These results indicate that a 134-basepair DNA fragment closely adjacent to the translation start site contains elements responsible for directing basal and stimulated expression of the EGF receptor gene.

FOOTNOTES

This work was supported by grants from the NIDDK (DK-13149) and the Council for Tobacco Research-U.S.A., Inc. (1622).

* Recipient of postdoctoral support from NIH Training Grant DK-07044.

Received for publication July 29, 1988. Accepted for publication November 15, 1988.




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