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Nucleic Acids Research, 1990, Vol. 18, No. 19 5677-5683
© 1990

MOLECULAR BIOLOGY

The promoter of the human parathyroid hormone gene contains a functional cyclic AMP-response element

Eva Rupp, Hubert Mayer and Edgar Wingender*

Gesellschaft für Biotechnologische Forschung mbH, Department of Genetics Mascheroder Weg 1, D-3300 Braunschweig, FRG

*To whom correspondence should be addressed

Received July 11, 1990. Revised September 3, 1990. Accepted September 3, 1990.

We have screened the sequence of the 394 base pairs upstream of the main transcriptional start site of the promoter of the human parathyroid hormone (PTH) gene for well-known protein recognition motifs with the aim to identify potential positive or negative regulatory elements. Within this region we found a potential cAMP-response element (CRE) besides several other putative binding sites for transcription factors. We fused promoter regions that contain this element and extend beyond the transcription start site to an appropriate reporter gene (CAT) and transfected different cell lines with these constructs. Translent expression of the CAT gene from these hybrid genes could be shown to be significantly stimulated by forskolin or isoproterenol thus proving the responsiveness of the whole promoter region towards elevated cAMP levels. DNase I protection studies revealed protein binding around the putative CRE (PTH-CRE) and an adjacent CCAAT element. Gel retardation assays with the PTH-CRE as well as the well-characterized CRE from the rat somatostatin promoter indicated specific binding of the same protein to both elements, although with a slightly reduced affinity of the PTH-CRE. Both of these elements were also able to confer cAMP-responsiveness to a heterologous promoter.


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