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

J. Sun, C. Oddoux, M. T. Gilbert, Y. Yan, A. Lazarus, W. G. Campbell, D. F. Catanzaro

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

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. Taken together, these data suggest that common cis-acting sequences direct renin gene expression to pituitary and placental cells and that the activity of these sequences is dependent on the presence of transcription factors specific to these cells. Our studies also indicate that 293 and JEG-3 cell lines may not be appropriate for the study of renin gene expression.

Original languageEnglish (US)
Pages (from-to)624-629
Number of pages6
JournalCirculation Research
Volume75
Issue number4
DOIs
StatePublished - Jan 1 1994

Fingerprint

Transcription Factor Pit-1
Primary Cell Culture
Renin
Binding Sites
Cell Line
DNA
Genes
5' Flanking Region
Gene Expression
Transcription Factors
Gels
TATA Box
Electrophoretic Mobility Shift Assay
Pituitary Gland
Adrenal Glands
Luciferases
Reporter Genes
Northern Blotting
Placenta

Keywords

  • pituitary
  • placenta
  • renin gene expression
  • transcription factor

ASJC Scopus subject areas

  • Physiology
  • Cardiology and Cardiovascular Medicine

Cite this

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. / Sun, J.; Oddoux, C.; Gilbert, M. T.; Yan, Y.; Lazarus, A.; Campbell, W. G.; Catanzaro, D. F.

In: Circulation Research, Vol. 75, No. 4, 01.01.1994, p. 624-629.

Research output: Contribution to journalArticle

@article{e6be1458d6f14938a4ccd8da1604cb65,
title = "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",
abstract = "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. Taken together, these data suggest that common cis-acting sequences direct renin gene expression to pituitary and placental cells and that the activity of these sequences is dependent on the presence of transcription factors specific to these cells. Our studies also indicate that 293 and JEG-3 cell lines may not be appropriate for the study of renin gene expression.",
keywords = "pituitary, placenta, renin gene expression, transcription factor",
author = "J. Sun and C. Oddoux and Gilbert, {M. T.} and Y. Yan and A. Lazarus and Campbell, {W. G.} and Catanzaro, {D. F.}",
year = "1994",
month = "1",
day = "1",
doi = "10.1161/01.RES.75.4.624",
language = "English (US)",
volume = "75",
pages = "624--629",
journal = "Circulation Research",
issn = "0009-7330",
publisher = "Lippincott Williams and Wilkins",
number = "4",

}

TY - JOUR

T1 - 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

AU - Sun, J.

AU - Oddoux, C.

AU - Gilbert, M. T.

AU - Yan, Y.

AU - Lazarus, A.

AU - Campbell, W. G.

AU - Catanzaro, D. F.

PY - 1994/1/1

Y1 - 1994/1/1

N2 - 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. Taken together, these data suggest that common cis-acting sequences direct renin gene expression to pituitary and placental cells and that the activity of these sequences is dependent on the presence of transcription factors specific to these cells. Our studies also indicate that 293 and JEG-3 cell lines may not be appropriate for the study of renin gene expression.

AB - 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. Taken together, these data suggest that common cis-acting sequences direct renin gene expression to pituitary and placental cells and that the activity of these sequences is dependent on the presence of transcription factors specific to these cells. Our studies also indicate that 293 and JEG-3 cell lines may not be appropriate for the study of renin gene expression.

KW - pituitary

KW - placenta

KW - renin gene expression

KW - transcription factor

UR - http://www.scopus.com/inward/record.url?scp=0028074809&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0028074809&partnerID=8YFLogxK

U2 - 10.1161/01.RES.75.4.624

DO - 10.1161/01.RES.75.4.624

M3 - Article

C2 - 7923608

AN - SCOPUS:0028074809

VL - 75

SP - 624

EP - 629

JO - Circulation Research

JF - Circulation Research

SN - 0009-7330

IS - 4

ER -