Determination and functional analysis of the consensus binding site for TFII-I family member BEN, implicated in Williams-Beuren syndrome.
The ubiquitously expressed TFII-I family of multifunctional transcription factors is involved in gene regulation as well as signaling. Despite the fact that they share significant sequence homology, these factors exhibit varied and distinct functions. The lack of knowledge about its binding sites and physiological target genes makes it more difficult to assign a definitive function for the TFII-I-related protein, BEN. We set out to determine its optimal binding site with the notion of predicting its physiological target genes. Here we report the identification of an optimal binding sequence for BEN by SELEX (systematic evolution of ligands by exponential enrichment) and confirm the relevance of this sequence by functional assays. We further performed a data base search to assign genes that have this consensus site(s) and validate several candidate genes by quantitative PCR upon stable silencing of BEN and by chromatin immunoprecipitation assay upon stable expression of BEN. Given that haploinsufficiency in BEN is causative to Williams-Beuren syndrome, these results may further lead to the identification of a set of physiologically relevant target genes for BEN and may help identify molecular determinants of Williams-Beuren syndrome.
Lazebnik
Maria B
MB
Programs in Genetics, Tufts University School of Medicine, Boston, Massachusetts 02111, USA.
Tussie-Luna
Maria Isabel
MI
Roy
Ananda L
AL
D000818:Animals; D001665:Binding Sites; D019556:COS Cells; D002522:Chlorocebus aethiops; D019295:Computational Biology / Q000379:methods; D005786:Gene Expression Regulation; D006801:Humans; D008024:Ligands; D051379:Mice; D008954:Models, Biological; D009124:Muscle Proteins / Q000378:metabolism*; D009687:Nuclear Proteins / Q000378:metabolism*; D011485:Protein Binding; D015534:Trans-Activators / Q000378:metabolism*; D014157:Transcription Factors / Q000378:metabolism*; D018980:Williams Syndrome / Q000378:metabolism*
C405493:GTF2IRD1 protein, human; C486558:Gtf2ird1 protein, mouse; D008024:Ligands; D009124:Muscle Proteins; D009687:Nuclear Proteins; D015534:Trans-Activators; D014157:Transcription Factors