Hormonal regulation of the bovine prolactin promoter in rat pituitary tumor cells.

S A Camper,Y A Yao,F M Rottman

doi:10.1016/s0021-9258(17)39017-8

Abstract

A 1-kilobase DNA fragment containing the promoter of the bovine prolactin gene was fused to the chloramphenicol acetyltransferase gene and the activity of the promoter was assayed by transfection of the fusion gene into COS-1, HeLa, and GH3 cells. Transcription of the chloramphenicol acetyltransferase gene driven by the prolactin promoter was detected only in GH3 cells, a rat pituitary tumor cell line. Epidermal growth factor and thyroid releasing hormone produced a stimulation of transcription, and the synthetic glucocorticoid hormone dexamethasone effected an inhibition of transcription from the prolactin promoter. None of these factors significantly affected transcription of the chloramphenicol acetyltransferase gene fused to the Rous sarcoma virus promoter. Deletion of all but 250 base pairs of bovine prolactin 5'-flanking DNA had no effect, indicating that the signals sufficient for both stimulation and inhibition of transcription reside in close proximity to the promoter.

Highlights

A 1-kilobaseDNA fragment containing the promotercomparison to include the human prolactin5’-flanking region of the bovine prolactin gene was fused to the chlor- (8),a highly conserved segment (89% homologous) spanning amphenicol acetyltransferasegene and theactivity of nucleotides -178 to -94 the promoterwas assayedby transfection of the fusionwas noted between the three species
Epidermal growth factor and thyroid releasing hormone producsetdimaulation of transcription, and thesynthetic glucocorticoid hormone dexamethasoneeffected an inhibitionof transcription fromtheprolactinpromoter.Noneof implicated as the location of regulatory functions in other eucaryotic genes, and is within a DNase I hypersensitive site mapped in theprolactin gene in rat pituitary cells (9)
Vaso- gene were monitored after treatment with EGF, thyrotropin releasing hormone (TRH), and active intestinal polypeptide probably acts through a CAMP- dexamethasone

Summary

Introduction

A 1-kilobaseDNA fragment containing the promotercomparison to include the human prolactin5’-flanking region of the bovine prolactin gene was fused to the chlor- (8),a highly conserved segment (89% homologous) spanning amphenicol acetyltransferasegene and theactivity of nucleotides -178 to -94 (numbering forthe bovine sequence) the promoterwas assayedby transfection of the fusionwas noted between the three species. Constructs containing the chloramphenicol acetyltransferase structural gene with SV40 splice site and polyadenylation signal driven by either the Rous sarcoma virus promoter (pRSVcat) or the SV40 early promoter (pSV2cat) (17, 20)were transfected as positive controls.

Results

Conclusion