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Endocrinology Vol. 139, No. 8 3423-3431
Copyright © 1998 by The Endocrine Society

ARTICLES

Characterization of a Negative Thyroid Hormone Response Element in the Rat Sodium, Potassium-Adenosine Triphosphatase {alpha}3 Gene Promoter1

Shing Chin, James Apriletti and Gregory Gick

Department of Biochemistry, State University of New York Health Science Center, Brooklyn, New York 11203; and Metabolic Research Unit, University of California (J.A.), San Francisco, California 94143

Address all correspondence and requests for reprints to: Gregory Gick, Ph.D., Department of Biochemistry, State University of New York Health Science Center, Brooklyn, New York 11203. E-mail: gickg11{at}hscbklyn.edu

The thyroid hormone L-T3 elicits either a stimulatory or an inhibitory effect on expression of the Na,K-adenosine triphosphatase {alpha}3-subunit gene in primary cultures of neonatal rat cardiac myocytes. The present study was undertaken to characterize a negative thyroid hormone response element present within the rat Na,K-adenosine triphosphatase {alpha}3-subunit gene proximal promoter. Transient transfection assays indicated that the DNA-binding domain of thyroid hormone receptor was essential for mediating repression of {alpha}3 gene transcription by thyroid hormone. This negative effect of thyroid hormone was enhanced in the presence of cotransfected retinoid X receptor and its ligand 9-cis-retinoic acid. Inhibition of {alpha}3 chimeric gene expression by thyroid hormone was dependent on the initial cell plating density. The negative thyroid hormone response element was localized to a region between nucleotides -68 to -6 of the {alpha}3 gene. Electrophoretic mobility shift assays showed that thyroid hormone receptor binds in a synergistic manner as a heterodimer with retinoid X receptor to two sites at positions -62 to -41 and -39 to -17 of the {alpha}3 gene promoter. The upstream and downstream heterodimer binding sites coexist with CAAT and TATA elements, respectively.




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Copyright © 1998 by The Endocrine Society