title: Two N-terminal self-association domains are required for the dominant negative transcriptional activity of WT1 Denys-Drash mutant proteins.
abstract: Patients with Denys-Drash syndrome (DDS) have been shown to be constitutionally heterozygous for mutations of the WT1 gene. Almost all DDS mutations inactivate or remove the DNA-binding zinc finger region of WT1 and the resulting mutant proteins appear to act in a dominant negative manner. This may occur via WT1 self-association, which has been shown to involve the first 180 amino acids. By creating a series of N-terminal deletions, we have further investigated WT1 self-association using a yeast di-hybrid system and an in vitro protein binding assay. Our results suggest that there are two distinct domains within the N-terminal region facilitating self-association, residing from amino acids 1-45 and 157-253. Co-transfection of WT1 with progressively shorter N-terminal constructs demonstrates that both of these sites are required for a dominant negative activity as assessed by activation of a reporter construct.
authors.affiliation: Center for Molecular and Cellular Biology, University of Queensland, St. Lucia, Australia.
authors.forename: G
authors.initials: G
authors.lastname: Holmes
authors.forename: S
authors.initials: S
authors.lastname: Boterashvili
authors.forename: M
authors.initials: M
authors.lastname: English
authors.forename: B
authors.initials: B
authors.lastname: Wainwright
authors.forename: J
authors.initials: J
authors.lastname: Licht
authors.forename: M
authors.initials: M
authors.lastname: Little
mesh_terms: D001483:Base Sequence; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D012734:Disorders of Sex Development / Q000235:genetics; D016162:Genes, Wilms Tumor; D006579:Heterozygote; D006801:Humans; D066298:In Vitro Techniques; D008297:Male; D015394:Molecular Structure; D009154:Mutation; D009838:Oligodeoxyribonucleotides / Q000235:genetics; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D015533:Transcriptional Activation; D014162:Transfection; D025721:WT1 Proteins
chemical_list: D004268:DNA-Binding Proteins; D009838:Oligodeoxyribonucleotides; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; D025721:WT1 Proteins