Overlapping RNA and DNA binding domains of the WT1 tumor suppressor gene product

  • Bardeesy N
  • Pelletier J
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The Wilms' tumour suppressor gene (wt1) is mutated in a subset of patients with Wilms' tumour and has a critical role in urogenital development. wt1 encodes a zinc finger transcription factor which regulates expression of several genes involved in cellular proliferation and differentiation. Although a number of studies have characterized the DNA binding properties of the WT1 protein, recent evidence has suggested that WT1 may also have a role in RNA metabolism. We have used an RNA selection method to identify WT1 binding ligands from a random RNA pool. Three groups of RNA ligands specifically recognized by WT1 were identified. Mutational analysis pinpointed ribonucleotide sequences critical for binding. Analysis of truncated WT1 proteins demonstrated that three of four zinc fingers were necessary for RNA-protein interaction. The naturally occurring WT1 isoforms with insertion of lysine, threonine and serine between zinc fingers three and four were unable to bind the selected RNAs. The selected RNA ligands competed with the cognate WT1 DNA binding site for complex formation with WT1. Our findings suggest potential cellular RNA target sequences for WT1 and provide tools for studying the structural and functional properties of this tumour suppressor protein.

Author-supplied keywords

  • *Genes, Suppressor, Tumor
  • Amino Acid Sequence
  • Base Sequence
  • Binding Sites
  • Consensus Sequence
  • DNA-Binding Proteins/chemistry/genetics/*metabolis
  • DNA/*chemistry/metabolism
  • Genes, Synthetic
  • Human
  • Kidney Neoplasms/*genetics
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • Nephroblastoma/*genetics
  • Oligodeoxyribonucleotides
  • RNA/*chemistry/metabolism
  • Recombinant Proteins/chemistry/metabolism
  • Support, Non-U.S. Gov't
  • Transcription Factors/chemistry/genetics/*metaboli
  • Zinc Fingers

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  • N Bardeesy

  • J Pelletier

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