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J Biol Chem, Vol. 273, Issue 33, 21145-21152, August 14, 1998
From the Smad proteins have been identified as mediators
of intracellular signal transduction by members of the transforming
growth factor-
Identification and Functional Characterization of a Smad Binding
Element (SBE) in the JunB Promoter That Acts as a
Transforming Growth Factor-
, Activin, and Bone Morphogenetic
Protein-inducible Enhancer
,
Department of Developmental Genetics,
Groningen Biomolecular Sciences and Biotechnology Institute, P. O. Box
14, 9750 AA Haren, The Netherlands and the ¶ Ludwig Institute for
Cancer Research, Box 595, Biomedical Center, S-75,
24 Uppsala, Sweden
(TGF-
) superfamily, which affect cell
proliferation, differentiation, as well as pattern formation during
early vertebrate development. Following receptor activation, Smads are
assembled into heteromeric complexes consisting of a pathway-restricted Smad and the common Smad4 that are subsequently translocated into the
nucleus where they are thought to play an important role in gene
transcription. Here we report the identification of Smad Binding
Elements (SBEs) composed of the sequence CAGACA in the promoter of the
JunB gene, an immediate early gene that is potently induced
by TGF-
, activin, and bone morphogenetic protein (BMP) 2. Two
JunB SBEs are arranged as an inverted repeat that is
transactivated in response to Smad3 and Smad4 co-overexpression and
shows inducible binding of a Smad3- and Smad4-containing complex in
nuclear extracts from TGF-
-treated cells. Bacterial-expressed Smad
proteins bind directly to the SBE. Multimerization of the SBE creates a
powerful TGF-
-inducible enhancer that is also responsive to activin
and BMPs. The identification of the sequence CAGACA as a direct binding site for Smad proteins will facilitate the identification of regulatory elements in genes that are activated by members of the TGF-
superfamily.
Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.
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