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