title: Flexible interaction of Drosophila Smad complexes with bipartite binding sites.
abstract: A subset of BMP-responsive enhancer elements are characterized by pairing of a GC-rich Smad1 binding site and an SBE-type Smad4 binding site. Such paired, or bipartite, sites are in some cases just 5 bp apart and thus might be contacted by a single Smad1-Smad4 complex. Other potential pairings are separated as much as 60 bp but it is not known whether such longer distances can be spanned by a Smad1-Smad4 complex, indeed binding of native Smad1-Smad4 complexes to any of these bipartite elements has yet to be reported. Here we report that a complex of the homologous Drosophila Smad proteins, Mad and Medea, is capable of concerted binding to GC-rich and SBE sites separated by as much as 20 bp. The wider the separation, the more severely binding affinity was reduced by shortening of the linker region that tethers the DNA binding domain of Medea. In contrast, length of the Mad linker did not affect the allowed distance between paired sites, rather it contributes specifically to Mad contact with the GC-rich site. Finally, we show that Smad1 and Smad4 can participate in binding to bipartite sites.
authors.affiliation: Laboratory of Genetics, University of Wisconsin, 425G Henry Mall, Madison, WI 53706, USA.
authors.forename: Sheng
authors.initials: S
authors.lastname: Gao
authors.forename: Allen
authors.initials: A
authors.lastname: Laughon
mesh_terms: D000818:Animals; D001665:Binding Sites / Q000502:physiology*; D004268:DNA-Binding Proteins / Q000502:physiology*; D029721:Drosophila Proteins / Q000502:physiology*; D011485:Protein Binding; D012097:Repressor Proteins / Q000502:physiology*; D051898:Smad1 Protein / Q000502:physiology*; D051901:Smad4 Protein / Q000502:physiology*; D014157:Transcription Factors / Q000502:physiology*
chemical_list: D004268:DNA-Binding Proteins; D029721:Drosophila Proteins; C112506:MAD protein, Drosophila; C112504:Med protein, Drosophila; D012097:Repressor Proteins; D051898:Smad1 Protein; D051901:Smad4 Protein; D014157:Transcription Factors; C118082:brk protein, Drosophila