Pituitary-specific transcription factor (Pit-1) binding site in the human renin gene 5'-flanking DNA stimulates promoter activity in placental cell primary cultures and pituitary lactosomatotropic cell lines.
Renin gene expression is limited to a number of specific tissues, including the kidney, adrenal glands, reproductive organs (of particular relevance to this study, the placenta), and the pituitary gland. In the present study, we investigated the human renin (hRen) 5'-flanking DNA sequences required to drive the expression of a luciferase reporter gene in placental and pituitary cells and in two cell lines, 293 and JEG-3, which have been proposed as model systems with which to study transcriptional regulation of renin genes. The activities of specific sequences in the hRen 5'-flanking DNA sequences in human placental cell primary cultures were very similar to those that we previously reported in pituitary cells, suggesting the involvement of common promoter elements and related transcription factors. Accordingly, the binding site for the pituitary-specific transcription factor (Pit-1) was the major determinant of renin promoter activity in both pituitary and placental cells. Gel mobility shift analysis showed a placental nuclear factor with a gel mobility different from that of Pit-1. However, Northern blot analysis failed to demonstrate abundant Pit-1-related mRNAs in renin-expressing cultures of chorionic and decidual cells, suggesting that the placental factor is not closely related to Pit-1. Although a factor from 293 cells also bound to the Pit-1 site, it had gel mobility shift characteristics different from Pit-1 and the placental factor. Moreover, the low promoter activity in 293 cells was independent of this site or, indeed, of sequences upstream from the TATA box. In JEG-3 cells, renin 5'-flanking DNA sequences showed virtually no transcriptional activity.(ABSTRACT TRUNCATED AT 250 WORDS)
Sun
J
J
Cardiovascular Center, Cornell University Medical College, New York, NY.
Oddoux
C
C
Gilbert
M T
MT
Yan
Y
Y
Lazarus
A
A
Campbell
W G
WG
Catanzaro
D F
DF
D001665:Binding Sites; D015152:Blotting, Northern; D002478:Cells, Cultured; D004247:DNA; D004268:DNA-Binding Proteins / Q000378:metabolism*; D015870:Gene Expression; D006801:Humans; D007150:Immunohistochemistry; D010902:Pituitary Gland / Q000166:cytology / Q000378:metabolism*; D010920:Placenta / Q000166:cytology / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D012083:Renin / Q000096:biosynthesis / Q000235:genetics*; D016385:TATA Box; D050817:Transcription Factor Pit-1; D014157:Transcription Factors / Q000378:metabolism*
D004268:DNA-Binding Proteins; C494187:POU1F1 protein, human; D050817:Transcription Factor Pit-1; D014157:Transcription Factors; D004247:DNA; D012083:Renin