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 language | English (US) |
|---|---|
| Pages (from-to) | 741-752 |
| Number of pages | 12 |
| Journal | Cell |
| Volume | 49 |
| Issue number | 6 |
| DOIs | |
| State | Published - Jun 19 1987 |
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All Science Journal Classification (ASJC) codes
- Biochemistry, Genetics and Molecular Biology(all)
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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 journal › Article
TY - JOUR
T1 - Purified transcription factor AP-1 interacts with TPA-inducible enhancer elements
AU - Lee, Wes
AU - Mitchell, Pamela
AU - Tjian, Robert
PY - 1987/6/19
Y1 - 1987/6/19
N2 - 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.
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.
UR - http://www.scopus.com/inward/record.url?scp=0023611441&partnerID=8YFLogxK
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U2 - 10.1016/0092-8674(87)90612-X
DO - 10.1016/0092-8674(87)90612-X
M3 - Article
C2 - 3034433
AN - SCOPUS:0023611441
VL - 49
SP - 741
EP - 752
JO - Cell
JF - Cell
SN - 0092-8674
IS - 6
ER -