The GC box is an important transcriptional regulatory element present in the promoters of many mammalian genes. In the present study we examine the effect of known GC-box-binding proteins on the promoter of the human elongation factor 1 A-1 (hEF1A-1) gene in human HeLa cells and Drosophila SL2 cells. In HeLa cells co-transfection with the GC-box-binding protein BTEB resulted in a 4–10-fold increase in hEF1A-1 promoter activity. This stimulation was dependent on a single GC box located between positions -69 and -50 of the promoter. Little or no effect was observed of other GC-box-binding proteins including Sp1, Sp3, Sp4 and BTEB2. In SL2 cells stimulation by Sp1 and Sp3 through the single GC box of the proximal promoter led to 13-fold and 21-fold increases respectively in promoter activity. Inclusion of further upstream sequences resulted in high levels of expression when Sp1 or Sp3 was co-transfected with the reporter plasmid. In this setting Sp1 stimulated transcription by 750-fold, whereas Sp3 was even more potent, yielding a 1150-fold stimulation. Mobility-shift assays performed with the promoter-proximal GC box demonstrated the binding of Sp1, Sp3 and Sp4 to this sequence. To our knowledge, the present study represents the first comparison of all known GC-box-binding proteins on a natural promoter.
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August 1998
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Research Article|
August 01 1998
Different Sp1 family members differentially affect transcription from the human elongation factor 1 A-1 gene promoter
S⊘ren J. NIELSEN;
1Department of Biostructural Chemistry, Institute of Molecular and Structural Biology, University of Aarhus, Gustav Wieds Vej 10C, Forskerparken, Århus, Denmark
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Morten PRÆSTEGAARD;
Morten PRÆSTEGAARD
1
1Department of Biostructural Chemistry, Institute of Molecular and Structural Biology, University of Aarhus, Gustav Wieds Vej 10C, Forskerparken, Århus, Denmark
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Helle F. J. ØRGENSEN;
Helle F. J. ØRGENSEN
1Department of Biostructural Chemistry, Institute of Molecular and Structural Biology, University of Aarhus, Gustav Wieds Vej 10C, Forskerparken, Århus, Denmark
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Brian F. C. CLARK
Brian F. C. CLARK
3
1Department of Biostructural Chemistry, Institute of Molecular and Structural Biology, University of Aarhus, Gustav Wieds Vej 10C, Forskerparken, Århus, Denmark
3To whom correspondence should be addressed (e-mail clark@biobase.dk).
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Publisher: Portland Press Ltd
Received:
February 12 1998
Revision Received:
April 02 1998
Accepted:
April 28 1998
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London © 1998
1998
Biochem J (1998) 333 (3): 511–517.
Article history
Received:
February 12 1998
Revision Received:
April 02 1998
Accepted:
April 28 1998
Citation
S⊘ren J. NIELSEN, Morten PRÆSTEGAARD, Helle F. J. ØRGENSEN, Brian F. C. CLARK; Different Sp1 family members differentially affect transcription from the human elongation factor 1 A-1 gene promoter. Biochem J 1 August 1998; 333 (3): 511–517. doi: https://doi.org/10.1042/bj3330511
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