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(Received for publication, March 22, 1995; and in revised form, May 12, 1995) From the Lactogenic hormone-dependent expression of the rat
Volume 270,
Number 30,
Issue of July 28, pp. 17962-17969, 1995
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.
and
Are Expressed in Mammary Epithelial
Cells and Bind to Multiple Sites in the
-Casein Gene Promoter
-casein
gene in mammary epithelial cells is controlled via a complex regulatory
region in the promoter. The sequence between -176 and -82
is the minimal region to confer the response to glucocorticoid hormone
and prolactin on a heterologous promoter. The response is further
enhanced by the region between -282 and -176. DNase I
footprinting experiments and electromobility shift assays revealed the
presence of four binding sites for CCAAT/enhancer-binding protein
(C/EBP) isoforms in the hormone response region between -220 and
-132. In nuclear extracts from mammary epithelial cells, the
prevalent C/EBP isoform binding to these sites is (C/EBP-).
C/EBP- is also present in mammary epithelial cells, whereas
C/EBP-
is not detectable. The C/EBP sites are located in close
proximity to the previously characterized binding sites for the
prolactin-inducible mammary gland factor/signal transducer and
activator of transcription-5, the nuclear factor YY1, and the
glucocorticoid receptor. The importance of the two proximal C/EBP
binding sites at the 5` border of the minimal region was tested by
mutational analysis. Mutations of each site were found to inhibit
strongly both the basal and the lactogenic hormone-induced
transcription of a -casein gene promoter chloramphenicol
acetyltransferase construct. The results implicate C/EBPs as important
regulators of -casein gene expression in the mammary epithelium.
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