Smad1 and Smad4 Are Components of the Bone Morphogenetic Protein-4 (BMP-4)-induced Transcription Complex of the Xvent-2B Promoter

To study the mechanism of transcriptional activation of the Xenopus homeobox geneXvent-2B, we have delineated the bone morphogenetic protein-4 (BMP-4)-responsive region between −275/−152 in the proximal promoter. Consistent with the BMP-4 inductive nature of this region, this element exhibits transc...

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Bibliographic Details
Published in:The Journal of biological chemistry 2000-07, Vol.275 (29), p.21827-21835
Main Authors: Henningfeld, Kristine A., Rastegar, Sepand, Adler, Guido, Knöchel, Walter
Format: Article
Language:English
Subjects:
Animals
Base Sequence
Bone Morphogenetic Protein 4
Bone Morphogenetic Proteins - genetics
DNA-Binding Proteins - genetics
Homeodomain Proteins - genetics
Molecular Sequence Data
Mutation
Nerve Growth Factors
Promoter Regions, Genetic
Smad Proteins
Smad1 protein
Smad4 Protein
Trans-Activators - genetics
Transcription Factors
Transcription, Genetic
Transcriptional Activation
Xenopus
Xenopus Proteins
Xvent-2B gene
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Summary:To study the mechanism of transcriptional activation of the Xenopus homeobox geneXvent-2B, we have delineated the bone morphogenetic protein-4 (BMP-4)-responsive region between −275/−152 in the proximal promoter. Consistent with the BMP-4 inductive nature of this region, this element exhibits transcriptional activation upon ectopic expression of Smad1 and Smad4. Electrophoretic mobility shift assays with total cellular extracts demonstrated that a DNA fragment encompassing this region was competent in the formation of a BMP-4-induced protein-DNA complex containing Smad1. Two different Smad binding regions were localized, a distal binding region for Smad1 containing two GCAT motifs and proximal AGNC binding sites for Smad4, the latter being conserved in other transforming growth factor-β-responsive elements. Mutation of the Smad4 binding motif completely abolished transcriptional activation, whereas mutation or deletion of the Smad1 recognition sequence inhibited Smad1/Smad4 responsiveness. These results provide a functional characterization and identification of a vertebrate Smad1/Smad4 DNA response element induced by BMP-4 signaling and offers insight into the transcriptional regulation of a component essential for dorsoventral patterning inXenopus embryos.
ISSN:0021-9258
1083-351X
DOI:10.1074/jbc.M000978200