Characterization of a Smad motif similar to Drosophila mad in the mouse Msx 1 promoter.
Mouse Msx 1 gene, orthologous of the Drosophila msh, is involved in several developmental processes. BMP family members are major proteins in the regulation of Msx 1 expression. BMP signaling activates Smad 1/5/8 proteins, which associate to Smad 4 before translocating to the nucleus. Analysis of Msx 1 promoter revealed the presence of three elements similar to the consensus established for Mad, the Smad 1 Drosophila counterpart. Notably, such an element was identified in an enhancer important for Msx 1 regulation. Gel shift analysis demonstrated that proteins from 13.5 dpc embryo associate to this enhancer. Remarkably, supershift assays showed that Smad proteins are present in the complex. Purified Smad 1 and 4 also bind to this fragment. We demonstrate that functional binding sites in this enhancer are confined to the Mad motif and flanking region. Our data suggest that this Mad motif may be functional in response to BMP signaling.
Alvarez Martinez
Cristina E
CE
Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil.
Binato
Renata
R
Gonzalez
Sayonara
S
Pereira
Monica
M
Robert
Benoit
B
Abdelhay
Eliana
E
D000818:Animals; D001483:Base Sequence; D016384:Consensus Sequence; D004268:DNA-Binding Proteins / Q000378:metabolism*; D029721:Drosophila Proteins; D024202:Electrophoretic Mobility Shift Assay; D004622:Embryo, Mammalian / Q000378:metabolism; D004742:Enhancer Elements, Genetic; D018398:Homeodomain Proteins / Q000235:genetics*; D051957:MSX1 Transcription Factor; D046911:Macromolecular Substances; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000378:metabolism; D011401:Promoter Regions, Genetic; D017384:Sequence Deletion; D051785:Smad Proteins; D051898:Smad1 Protein; D051901:Smad4 Protein; D015534:Trans-Activators / Q000378:metabolism*; D014157:Transcription Factors
D004268:DNA-Binding Proteins; D029721:Drosophila Proteins; D018398:Homeodomain Proteins; C112506:MAD protein, Drosophila; D051957:MSX1 Transcription Factor; D046911:Macromolecular Substances; D009841:Oligonucleotides; D051785:Smad Proteins; D051898:Smad1 Protein; C497401:Smad1 protein, mouse; D051901:Smad4 Protein; C490810:Smad4 protein, mouse; D015534:Trans-Activators; D014157:Transcription Factors