Nucleic Acids Research, 1989, Vol. 17, No. 7 2835-2848
© 1989
MOLECULAR BIOLOGY |
Glucocorticoid and estrogen regulation of a rat T-kininogen gene
Department of Molecular Genetics, Biochemistry and Microbiology, University of Cincinnati Cincinnati, OH 45267, USA
*To whom correspondence should be addressed
Received September 26, 1988. Revised February 16, 1989. Accepted February 16, 1989.
We have examined the regulation of a rat T-kininogen gene by glucocortieoid and estrogen. Expression of the endogenous gene in a rat hepatoma cell line is increased 5-fold and 2-fold in response to dexamethasone and 17 ß-estradiol-3-benzoate, respectively. Various deletion constructs of the 5' region of an isolated T-kininogen gene were fused to a ehloramphenicol acetyltransferase (CAT) gene and introduced into the hepatoma cells by electroporation. Analysis of the CAT activity in cell extracts after treatment with glucocorticoid or estrogen revealed that a fragment from –167 to +52 is sufficient to confer full induction. An additional deletion in this region was unresponsive, while a larger fragment (–612 to –100) linked to a heterologous promoter did result in regulated expression. These results suggested that the sequence responsible for the hormonal response was located at –167 to –100 from the transcription start site. This 67 bp region contains a consensus for the core sequence of the glucocorticoid responsive element (GRE) and the estrogen responsive element (ERE). Interestingly these elements are located within 7 bp of each other and both sequences overlap a 16 bp palindrome that may be important in hormone receptor-DNA recognition.
+Present adrress: Greenwood Genetic Center, Greenwood, SC 29646, USA