Identification of an estrogen response element in the 3′-flanking region of the murine c-fos protooncogene

Salman M. Hyder, George M. Stancel, Zafar Nawaz, Donald P. McDonnell, David S. Loose-Mitchell

Research output: Contribution to journalArticle

96 Citations (Scopus)

Abstract

We have used transient transfection assays with reporter plasmids expressing chloramphenicol acetyltransferase, linked to regions of mouse c-fos, to identify a specific estrogen response element (ERE) in this protooncogene. This element is located in the untranslated 3′-flanking region of the c-fos gene, 5 kilobases (kb) downstream from the c-fos promoter and 1.5 kb downstream of the poly(A) signal. This element confers estrogen responsiveness to chloramphenicol acetyltransferase reporters linked to both the herpes simplex virus thymidine kinase promoter and the homologous c-fos promoter. Deletion analysis localized the response element to a 200-base pair fragment which contains the element GGTCACCACAGCC that resembles the consensus ERE sequence GGTCACAGTGACC originally identified in Xenopus vitellogenin A2 gene. A synthetic 36-base pair oligodeoxynucleotide containing this c-fos sequence conferred estrogen inducibility to the thymidine kinase promoter. The corresponding sequence also induced reporter activity when present in the c-fos gene fragment 3 kb from the thymidine kinase promoter. Gel-shift experiments demonstrated that synthetic oligonucleotides containing either the consensus ERE or the c-fos element bind human estrogen receptor obtained from a yeast expression system. However, the mobility of the shifted band is faster for the fos-ERE-complex than the consensus ERE complex suggesting that the three-dimensional structure of the protein-DNA complexes is different or that other factors are differentially involved in the two reactions. When the 5′-GGTCA sequence present in the c-fos ERE is mutated to 5′-TTTCA, transcriptional activation and receptor binding activities are both lost. Mutation of the CAGCC-3′ element corresponding to the second half-site of the c-fos sequence also led to the loss of receptor binding activity, suggesting that both half-sites of this element are involved in this function. The estrogen induction mediated by either the c-fos or the consensus ERE was blunted by the antiestrogen tamoxifen. Based on these studies, we believe the 3′-fos ERE sequence we have identified may be a major cis-acting element involved in the physiological regulation of the gene by estrogens in vivo.

Original languageEnglish
Pages (from-to)18047-18054
Number of pages8
JournalJournal of Biological Chemistry
Volume267
Issue number25
StatePublished - Sep 5 1992
Externally publishedYes

Fingerprint

3' Flanking Region
Response Elements
Estrogens
Thymidine Kinase
Genes
fos Genes
Chloramphenicol O-Acetyltransferase
Base Pairing
Vitellogenins
Poly A
Estrogen Receptor Modulators
Oligodeoxyribonucleotides
Simplexvirus
Tamoxifen
Xenopus
Viruses
varespladib methyl
Oligonucleotides
Estrogen Receptors
Yeast

ASJC Scopus subject areas

  • Biochemistry

Cite this

Hyder, S. M., Stancel, G. M., Nawaz, Z., McDonnell, D. P., & Loose-Mitchell, D. S. (1992). Identification of an estrogen response element in the 3′-flanking region of the murine c-fos protooncogene. Journal of Biological Chemistry, 267(25), 18047-18054.

Identification of an estrogen response element in the 3′-flanking region of the murine c-fos protooncogene. / Hyder, Salman M.; Stancel, George M.; Nawaz, Zafar; McDonnell, Donald P.; Loose-Mitchell, David S.

In: Journal of Biological Chemistry, Vol. 267, No. 25, 05.09.1992, p. 18047-18054.

Research output: Contribution to journalArticle

Hyder, SM, Stancel, GM, Nawaz, Z, McDonnell, DP & Loose-Mitchell, DS 1992, 'Identification of an estrogen response element in the 3′-flanking region of the murine c-fos protooncogene', Journal of Biological Chemistry, vol. 267, no. 25, pp. 18047-18054.
Hyder, Salman M. ; Stancel, George M. ; Nawaz, Zafar ; McDonnell, Donald P. ; Loose-Mitchell, David S. / Identification of an estrogen response element in the 3′-flanking region of the murine c-fos protooncogene. In: Journal of Biological Chemistry. 1992 ; Vol. 267, No. 25. pp. 18047-18054.
@article{39a98e20e619436384e71abe3b5ed1fa,
title = "Identification of an estrogen response element in the 3′-flanking region of the murine c-fos protooncogene",
abstract = "We have used transient transfection assays with reporter plasmids expressing chloramphenicol acetyltransferase, linked to regions of mouse c-fos, to identify a specific estrogen response element (ERE) in this protooncogene. This element is located in the untranslated 3′-flanking region of the c-fos gene, 5 kilobases (kb) downstream from the c-fos promoter and 1.5 kb downstream of the poly(A) signal. This element confers estrogen responsiveness to chloramphenicol acetyltransferase reporters linked to both the herpes simplex virus thymidine kinase promoter and the homologous c-fos promoter. Deletion analysis localized the response element to a 200-base pair fragment which contains the element GGTCACCACAGCC that resembles the consensus ERE sequence GGTCACAGTGACC originally identified in Xenopus vitellogenin A2 gene. A synthetic 36-base pair oligodeoxynucleotide containing this c-fos sequence conferred estrogen inducibility to the thymidine kinase promoter. The corresponding sequence also induced reporter activity when present in the c-fos gene fragment 3 kb from the thymidine kinase promoter. Gel-shift experiments demonstrated that synthetic oligonucleotides containing either the consensus ERE or the c-fos element bind human estrogen receptor obtained from a yeast expression system. However, the mobility of the shifted band is faster for the fos-ERE-complex than the consensus ERE complex suggesting that the three-dimensional structure of the protein-DNA complexes is different or that other factors are differentially involved in the two reactions. When the 5′-GGTCA sequence present in the c-fos ERE is mutated to 5′-TTTCA, transcriptional activation and receptor binding activities are both lost. Mutation of the CAGCC-3′ element corresponding to the second half-site of the c-fos sequence also led to the loss of receptor binding activity, suggesting that both half-sites of this element are involved in this function. The estrogen induction mediated by either the c-fos or the consensus ERE was blunted by the antiestrogen tamoxifen. Based on these studies, we believe the 3′-fos ERE sequence we have identified may be a major cis-acting element involved in the physiological regulation of the gene by estrogens in vivo.",
author = "Hyder, {Salman M.} and Stancel, {George M.} and Zafar Nawaz and McDonnell, {Donald P.} and Loose-Mitchell, {David S.}",
year = "1992",
month = "9",
day = "5",
language = "English",
volume = "267",
pages = "18047--18054",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "25",

}

TY - JOUR

T1 - Identification of an estrogen response element in the 3′-flanking region of the murine c-fos protooncogene

AU - Hyder, Salman M.

AU - Stancel, George M.

AU - Nawaz, Zafar

AU - McDonnell, Donald P.

AU - Loose-Mitchell, David S.

PY - 1992/9/5

Y1 - 1992/9/5

N2 - We have used transient transfection assays with reporter plasmids expressing chloramphenicol acetyltransferase, linked to regions of mouse c-fos, to identify a specific estrogen response element (ERE) in this protooncogene. This element is located in the untranslated 3′-flanking region of the c-fos gene, 5 kilobases (kb) downstream from the c-fos promoter and 1.5 kb downstream of the poly(A) signal. This element confers estrogen responsiveness to chloramphenicol acetyltransferase reporters linked to both the herpes simplex virus thymidine kinase promoter and the homologous c-fos promoter. Deletion analysis localized the response element to a 200-base pair fragment which contains the element GGTCACCACAGCC that resembles the consensus ERE sequence GGTCACAGTGACC originally identified in Xenopus vitellogenin A2 gene. A synthetic 36-base pair oligodeoxynucleotide containing this c-fos sequence conferred estrogen inducibility to the thymidine kinase promoter. The corresponding sequence also induced reporter activity when present in the c-fos gene fragment 3 kb from the thymidine kinase promoter. Gel-shift experiments demonstrated that synthetic oligonucleotides containing either the consensus ERE or the c-fos element bind human estrogen receptor obtained from a yeast expression system. However, the mobility of the shifted band is faster for the fos-ERE-complex than the consensus ERE complex suggesting that the three-dimensional structure of the protein-DNA complexes is different or that other factors are differentially involved in the two reactions. When the 5′-GGTCA sequence present in the c-fos ERE is mutated to 5′-TTTCA, transcriptional activation and receptor binding activities are both lost. Mutation of the CAGCC-3′ element corresponding to the second half-site of the c-fos sequence also led to the loss of receptor binding activity, suggesting that both half-sites of this element are involved in this function. The estrogen induction mediated by either the c-fos or the consensus ERE was blunted by the antiestrogen tamoxifen. Based on these studies, we believe the 3′-fos ERE sequence we have identified may be a major cis-acting element involved in the physiological regulation of the gene by estrogens in vivo.

AB - We have used transient transfection assays with reporter plasmids expressing chloramphenicol acetyltransferase, linked to regions of mouse c-fos, to identify a specific estrogen response element (ERE) in this protooncogene. This element is located in the untranslated 3′-flanking region of the c-fos gene, 5 kilobases (kb) downstream from the c-fos promoter and 1.5 kb downstream of the poly(A) signal. This element confers estrogen responsiveness to chloramphenicol acetyltransferase reporters linked to both the herpes simplex virus thymidine kinase promoter and the homologous c-fos promoter. Deletion analysis localized the response element to a 200-base pair fragment which contains the element GGTCACCACAGCC that resembles the consensus ERE sequence GGTCACAGTGACC originally identified in Xenopus vitellogenin A2 gene. A synthetic 36-base pair oligodeoxynucleotide containing this c-fos sequence conferred estrogen inducibility to the thymidine kinase promoter. The corresponding sequence also induced reporter activity when present in the c-fos gene fragment 3 kb from the thymidine kinase promoter. Gel-shift experiments demonstrated that synthetic oligonucleotides containing either the consensus ERE or the c-fos element bind human estrogen receptor obtained from a yeast expression system. However, the mobility of the shifted band is faster for the fos-ERE-complex than the consensus ERE complex suggesting that the three-dimensional structure of the protein-DNA complexes is different or that other factors are differentially involved in the two reactions. When the 5′-GGTCA sequence present in the c-fos ERE is mutated to 5′-TTTCA, transcriptional activation and receptor binding activities are both lost. Mutation of the CAGCC-3′ element corresponding to the second half-site of the c-fos sequence also led to the loss of receptor binding activity, suggesting that both half-sites of this element are involved in this function. The estrogen induction mediated by either the c-fos or the consensus ERE was blunted by the antiestrogen tamoxifen. Based on these studies, we believe the 3′-fos ERE sequence we have identified may be a major cis-acting element involved in the physiological regulation of the gene by estrogens in vivo.

UR - http://www.scopus.com/inward/record.url?scp=0026744196&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0026744196&partnerID=8YFLogxK

M3 - Article

C2 - 1517237

AN - SCOPUS:0026744196

VL - 267

SP - 18047

EP - 18054

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 25

ER -