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Originally published In Press as doi:10.1074/jbc.M110225200 on January 3, 2002

J. Biol. Chem., Vol. 277, Issue 10, 8061-8067, March 8, 2002
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BRCA1 Regulates GADD45 through Its Interactions with the OCT-1 and CAAT Motifs*

Wenhong FanDagger , Shunqian JinDagger , Tong TongDagger , Hongcheng ZhaoDagger , Feiyue FanDagger , Michael J. AntinoreDagger , Baskaran Rajasekaran§, Min Wu, and Qimin ZhanDagger §||

From the Dagger  Department of Radiation Oncology, Cancer Institute, and § Department of Molecular Genetics and Biochemistry, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15213 and  National Laboratory of Molecular Oncology, Cancer Institute, Chinese Academy of Medical Sciences, Beijing 100021, China

BRCA1, a breast and ovarian cancer susceptibility gene, has been implicated in gene regulation. Previous studies demonstrate that BRCA1 induces GADD45, a p53-regulated and stress-inducible gene that plays an important role in cellular response to DNA damage. However, the mechanism(s) by which BRCA1 regulates GADD45 remains unclear. In this report, we have shown that BRCA1 activation of the GADD45 promoter is mediated through the OCT-1 and CAAT motifs located at the GADD45 promoter region. Site-directed mutations of both OCT-1 and CAAT motifs abrogate induction of the GADD45 promoter by BRCA1. Both OCT-1 and CAAT motifs are able to confer BRCA1 inducibility in a non-related minimal promoter. Physical associations of BRCA1 protein with transcription factors Oct-1 and NF-YA, which directly bind to the OCT-1 and CAAT motifs, are established by biotin-streptavidin pull-down and coimmunoprecipitation assays. Such protein interactions are required for interaction of BRCA1 with the GADD45 promoter because either immunodepletion of Oct-1 and NF-YA proteins or mutations in the OCT-1 and CAAT motifs disrupt BRCA1 binding to the GADD45 promoter. These findings indicate that BRCA1 can up-regulate its targeted genes through protein-protein interactions and provide a novel mechanism by which BRCA1 participates in transcriptional regulation.

* This work was supported in part by National Institutes of Health Grant R01 CA 93640-01 and Department of Defense Grant DAMD 17-00-1-0414.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.

|| To whom correspondence should be addressed: Cancer Institute, University of Pittsburgh School of Medicine, BST W-945, 200 Lothrop St., Pittsburgh, PA 15213. Fax: 412624-0295; E-mail: Qzhan@pitt.edu.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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