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Originally published In Press as doi:10.1074/jbc.M011643200 on February 13, 2001

J. Biol. Chem., Vol. 276, Issue 19, 15598-15608, May 11, 2001
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The Tumor Suppressor Protein p53 Requires a Cofactor to Activate Transcriptionally the Human BAX Promoter*

Edward C. Thornborrow and James J. ManfrediDagger

From The Derald H. Ruttenberg Cancer Center, Mount Sinai School of Medicine, New York, New York 10029

An important regulator of the proapoptotic BAX is the tumor suppressor protein p53. Unlike the p21 gene, in which p53-dependent transcriptional activation is mediated by a response element containing two consensus p53 half-sites, it previously was reported that activation of the BAX element by p53 requires additional sequences. Here, it is demonstrated that the minimal BAX response element capable of mediating p53-dependent transcriptional activation consists of two p53 half-sites plus an adjacent 6 base pairs (5'-GGGCGT-3'). This GC-rich region constitutes a "GC box" capable both of binding members of the Sp family of transcription factors, including Sp1 in vitro, and of conferring Sp1-dependent transcriptional activation on a minimal promoter in cells. Mutations within this GC box abrogated the ability of p53 to activate transcription without affecting the affinity of p53 for its binding site, demonstrating that these 6 bases are required for p53-dependent activation. In addition, a positive correlation was observed between the ability of p53 to activate transcription in cells and the ability of Sp1 to bind this response element in vitro. Mutations that inhibited Sp1 binding also blocked the ability of p53 to activate transcription through this element. Together, these results suggest a model in which p53 requires the cooperation of Sp1 or a Sp1-like factor to mediate transcriptional activation of the human BAX promoter.

* This work was supported by NCI Grant CA69161 from the National Institutes of Health and the Breast Cancer Program of the United States Army Medical Research and Materiel Command Grants DAMD-17-97-1-7336, DAMD-17-97-1-7337, and DAMD-17-99-1-9308).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Dagger To whom correspondence should be addressed: Cancer Center, Box 1130, Mount Sinai School of Medicine, New York, NY 10029. Tel.: 212-659-5495; Fax: 212-849-2446.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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