Purified transcription factor AP-1 interacts with TPA-inducible enhancer elements

Wes Lee, Pamela Mitchell, Robert Tjian

Research output: Contribution to journalArticle

1330 Citations (Scopus)

Abstract

The enhancer-binding protein AP-1 has been purified to >95% homogeneity from HeLa cells by sequence-specific DNA affinity chromatography and identified as a 47 kd polypeptide. Purified AP-1 activates transcription in vitro of the wild-type human metallothionein IIA (hMT IIA) gene but not mutant hMT IIA promoters lacking AP-1 recognition sites. DNAase I protection analysis indicates that genetically defined enhancer elements in hMT IIA, SV40, and the human collagenase gene contain high-affinity AP-1-binding sites, each with a conserved recognition motif, TGACTCA. These three genes are transcriptionally induced by treatment of cells with the tumor promoter TPA. Here we demonstrate that multiple synthetic copies of the consensus AP-1-binding site can act as TPA-inducible enhancers in various plasmid constructs after transfection into HeLa cells. These findings suggest that AP-1 is a transcription factor that functions by interacting with a specific enhancer element, and that its activities may be modulated by treatment of cells with TPA, known to stimulate protein kinase C.

Original languageEnglish (US)
Pages (from-to)741-752
Number of pages12
JournalCell
Volume49
Issue number6
DOIs
StatePublished - Jun 19 1987

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Transcription Factor AP-1
Metallothionein
Genes
HeLa Cells
Binding Sites
Cells
Affinity chromatography
Deoxyribonuclease I
Collagenases
Transcription
Affinity Chromatography
Carcinogens
Protein Kinase C
Transfection
Plasmids
Transcription Factors
Peptides
DNA

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Lee, Wes ; Mitchell, Pamela ; Tjian, Robert. / Purified transcription factor AP-1 interacts with TPA-inducible enhancer elements. In: Cell. 1987 ; Vol. 49, No. 6. pp. 741-752.
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Purified transcription factor AP-1 interacts with TPA-inducible enhancer elements. / Lee, Wes; Mitchell, Pamela; Tjian, Robert.

In: Cell, Vol. 49, No. 6, 19.06.1987, p. 741-752.

Research output: Contribution to journalArticle

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AB - The enhancer-binding protein AP-1 has been purified to >95% homogeneity from HeLa cells by sequence-specific DNA affinity chromatography and identified as a 47 kd polypeptide. Purified AP-1 activates transcription in vitro of the wild-type human metallothionein IIA (hMT IIA) gene but not mutant hMT IIA promoters lacking AP-1 recognition sites. DNAase I protection analysis indicates that genetically defined enhancer elements in hMT IIA, SV40, and the human collagenase gene contain high-affinity AP-1-binding sites, each with a conserved recognition motif, TGACTCA. These three genes are transcriptionally induced by treatment of cells with the tumor promoter TPA. Here we demonstrate that multiple synthetic copies of the consensus AP-1-binding site can act as TPA-inducible enhancers in various plasmid constructs after transfection into HeLa cells. These findings suggest that AP-1 is a transcription factor that functions by interacting with a specific enhancer element, and that its activities may be modulated by treatment of cells with TPA, known to stimulate protein kinase C.

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