Direct binding of Smad1 and Smad4 to two distinct motifs mediates bone morphogenetic protein-specific transcriptional activation of Id1 gene Journal Article
| Authors: | López-Rovira, T.; Chalaux, E.; Massagué, J.; Rosa, J. L.; Ventura, F. |
| Article Title: | Direct binding of Smad1 and Smad4 to two distinct motifs mediates bone morphogenetic protein-specific transcriptional activation of Id1 gene |
| Abstract: | Bone morphogenetic proteins (BMPs) are potent inhibitors of myoblast differentiation and inducers of bone formation both in vivo and in vitro. Expression of Idl, a negative regulator of basic helix-loop-helix transcription factors, is up-regulated by BMPs and contributes to the antimyogenic effects of this family of cytokines. In this report, we have identified a specific BMP-2 immediate early response enhancer in the human Id1 gene. Transcriptional activation of the enhancer was increased by overexpression of BMP-responsive Smads, and Smad4 and was completely abrogated in Smad4-deficient cells. Deletion analysis demonstrates that the responsive region is composed of two separate DNA binding elements, a set of overlapping GC boxes, which bind BMP-regulated Smads upon BMP stimulation, and three repeats of CAGAC boxes. Gel shift and oligonucleotide pull-down assays demonstrated that these two types of motifs were capable of binding their corresponding Smads. However, deletion or mutation of either DNA binding element was nonadditive, since disruption of either GC or CAGAC boxes resulted in complete or severe loss of BMP-2 responsiveness. These data suggest the simultaneous requirement of two independent DNA binding elements to allow functional cooperativity of BMP-regulated Smads and Smad4 in BMP-activated gene promoters. |
| Keywords: | signal transduction; unclassified drug; dna binding protein; human cell; promoter region; dna-binding proteins; proteins; animals; gene overexpression; genes; bone morphogenetic protein; smad protein; smad1 protein; transforming growth factor beta; protein dna binding; transcription initiation; cell line; protein binding; transfection; mutational analysis; transcription factors; gene expression regulation; cloning, molecular; gene activation; dna; kinetics; gene disruption; recombinant proteins; bone; smad proteins; dna sequence; binding sites; genes, reporter; trans-activators; inhibitor of differentiation protein 1; luciferases; biochemistry; repressor proteins; mutations; bone morphogenetic proteins; enhancer region; cells; smad4 protein; promoter regions (genetics); trans-activation (genetics); helix-loop-helix motifs; human; priority journal; article |
| Journal Title: | Journal of Biological Chemistry |
| Volume: | 277 |
| Issue: | 5 |
| ISSN: | 0021-9258 |
| Publisher: | American Society for Biochemistry and Molecular Biology |
| Date Published: | 2002-02-01 |
| Start Page: | 3176 |
| End Page: | 3185 |
| Language: | English |
| DOI: | 10.1074/jbc.M106826200 |
| PUBMED: | 11700304 |
| PROVIDER: | scopus |
| DOI/URL: | |
| Notes: | Export Date: 14 November 2014 -- Source: Scopus |
