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The WT1 tumor suppressor gene encodes four isoforms of
a zinc finger transcription factor with both activation and repression
functions which are dependent upon promoter architecture. Using a
simple HSV- tk promoter containing 5`-Egr-1/WT1-binding sites,
we found that WT1 isoforms (A) and (B) strongly activated
transcription. WT1(A) and (B) bound equally well to the
Egr-1/WT1-binding site, but WT1(B), which contains a 17 amino acid
insertion compared to WT1(A), was a consistently stronger activator of
transcription than WT1(A). Transcriptional activation by wild-type WT1
was inhibited by coexpression of WT(PM) or WT(AR),
genetically defined dominant negative alleles of WT1. In
vitro, as well as in the yeast two-hybrid system, WT1 protein
associated with itself and with dominant negative mutant proteins. The
major domain required for self-association and inhibition of
transcriptional activation mapped to the first 182 amino acids of WT1.
Dominant negative WT1 alleles may play a role in tumorigenesis by
associating with wild-type WT1 proteins and decreasing their
transcriptional activity.
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