We analysed in detail the minimal promoter of transcription factor Sp1, which extends 217bp from the initiation of transcription. Within this sequence we identified putative binding sites for Sp1, nuclear factor Y (NF-Y), activator protein 2 ('AP-2'), CCAAT/enhancer-binding protein ('C/EBP') and E2F transcription factors. In one case, the boxes for Sp1 and NF-Y are overlapping. Gel-shift and supershift assays demonstrated specific binding of Sp1, Sp3 and NF-Y proteins. Transient transfections and luciferase assays revealed activation of the Sp1 minimal promoter upon overexpression of Sp1 itself, NF-Y and E2F. Whereas overexpression of NF-Y or E2F had an additive effect on Sp1 overexpression, the activation of Sp1 transcription due to Sp1 was counteracted by Sp3 overexpression. Mutagenesis analysis of the NFY/Sp1-overlapping box revealed that both factors compete for this box, and that when the NF-Y site of this overlapping box is specifically mutated there is an increase in Sp1 binding, thus increasing transcriptional activity. These results help to explain the complex regulation of the Sp1 gene, which depends on the relative amounts of Sp1, Sp3, E2F and NF-Y proteins in the cell.
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April 2003
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Research Article|
April 15 2003
Transcriptional regulation of the human Sp1 gene promoter by the specificity protein (Sp) family members nuclear factor Y (NF-Y) and E2F
Marta NICOLÁS;
Marta NICOLÁS
Department of Biochemistry and Molecular Biology, School of Pharmacy, University of Barcelona, E-08028 Barcelona, Spain
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Vèronique NOÉ;
Vèronique NOÉ
Department of Biochemistry and Molecular Biology, School of Pharmacy, University of Barcelona, E-08028 Barcelona, Spain
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Carlos J. CIUDAD
Carlos J. CIUDAD
1
Department of Biochemistry and Molecular Biology, School of Pharmacy, University of Barcelona, E-08028 Barcelona, Spain
1To whom correspondence should be addressed (e-mail Cciudad@farmacia.far.ub.es).
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Publisher: Portland Press Ltd
Received:
July 25 2002
Revision Received:
December 05 2002
Accepted:
January 03 2003
Accepted Manuscript online:
January 03 2003
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London ©2003
2003
Biochem J (2003) 371 (2): 265–275.
Article history
Received:
July 25 2002
Revision Received:
December 05 2002
Accepted:
January 03 2003
Accepted Manuscript online:
January 03 2003
Citation
Marta NICOLÁS, Vèronique NOÉ, Carlos J. CIUDAD; Transcriptional regulation of the human Sp1 gene promoter by the specificity protein (Sp) family members nuclear factor Y (NF-Y) and E2F. Biochem J 15 April 2003; 371 (2): 265–275. doi: https://doi.org/10.1042/bj20021166
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