Binding of the Wilms' Tumor Locus Zinc Finger Protein to the EGR-1 Consensus Sequence

The Wilms' tumor locus (WTL) at 11p13 contains a gene that encodes a zinc finger-containing protein that has characteristics of a DNA-binding protein. However, binding of this protein to DNA in a sequence-specific manner has not been demonstrated. A synthetic gene was constructed that contained...

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Bibliographic Details
Published in:Science (American Association for the Advancement of Science) 1990-11, Vol.250 (4985), p.1259-1262
Main Authors: Rauscher, Frank J., Morris, Jennifer F., Tournay, Olivia E., Cook, Donna M., Curran, Tom
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Base Sequence
Binding Sites
Binding, Competitive
Carcinogenesis
Chromosomes, Human, Pair 11
Consensus Sequence
Deoxyribonucleic acid
DNA
DNA - genetics
DNA - metabolism
DNA binding proteins
DNA probes
DNA-Binding Proteins - genetics
DNA-Binding Proteins - metabolism
Early Growth Response Protein 1
Escherichia coli - genetics
Gels
Genes, Wilms Tumor
Genetic aspects
Genetic mutation
Genetics
Humans
Immediate-Early Proteins
Molecular Sequence Data
Mutation
Nephroblastoma
Oligonucleotide probes
Oligonucleotides
Oligonucleotides - metabolism
Polymerase Chain Reaction
Proteins
Recombinant Proteins - metabolism
Restriction Mapping
Transcription Factors - genetics
Transcription Factors - metabolism
Tumors
Zinc
Zinc Fingers - genetics
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Description
Summary:The Wilms' tumor locus (WTL) at 11p13 contains a gene that encodes a zinc finger-containing protein that has characteristics of a DNA-binding protein. However, binding of this protein to DNA in a sequence-specific manner has not been demonstrated. A synthetic gene was constructed that contained the zinc finger region, and the protein was expressed in Escherichia coli. The recombinant protein was used to identify a specific DNA binding site from a pool of degenerate oligonucleotides. The binding sites obtained were similar to the sequence recognized by the early growth response-1 (EGR-1) gene product, a zinc finger-containing protein that is induced by mitogenic stimuli. A mutation in the zinc finger region of the protein originally identified in a Wilms' tumor patient abolished its DNA-binding activity. These results suggest that the WTL protein may act at the DNA binding site of a growth factor-inducible gene and that loss of DNA-binding activity contributes to the tumorigenic process.
ISSN:0036-8075
1095-9203
DOI:10.1126/science.2244209