Tumor-associated WT1 missense mutants indicate that transcriptional activation by WT1 is critical for growth control.
The WT1 gene encodes a zinc finger DNA binding transcription factor and is mutated in up to 15% of Wilms tumor cases. The WT1 protein binds to the promoters of many genes through GC- or TC-rich sequences and can function both as a transcriptional repressor and an activator in co-transfection assays depending on the cell type, the structure of the test promoter, and even the expression vectors used. Engineered expression of WT1 can lead to growth suppression by both cell cycle arrest and induction of apoptosis. However, the transcriptional activity of WT1 that is required for growth control was not defined. We found that three N-terminal tumor-associated missense mutations of WT1 were defective for activation of both a synthetic reporter containing WT1-binding sites as well as the promoter of a WT1 responsive gene, p21. These mutants failed to inhibit cell growth but still retain their ability to repress several putative WT1 target promoters. These results indicate that activation and not repression by WT1 is the critical transcriptional activity of the protein responsible for its growth suppressing properties.
English
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Derald H. Ruttenberg Cancer Center and the Department of Medicine, Mount Sinai School of Medicine, New York, New York 10029, USA.
Licht
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D016475:3T3 Cells; D000818:Animals; D001483:Base Sequence; D002455:Cell Division / Q000235:genetics*; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins / Q000235:genetics; D017931:DNA Primers; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D017930:Genes, Reporter; D016162:Genes, Wilms Tumor; D006801:Humans; D051379:Mice; D020125:Mutation, Missense; D009369:Neoplasms / Q000235:genetics / Q000473:pathology*; D011401:Promoter Regions, Genetic; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D015533:Transcriptional Activation; D025721:WT1 Proteins; D016335:Zinc Fingers
C493956:CDKN1A protein, human; C493957:Cdkn1a protein, mouse; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins; D017931:DNA Primers; D004268:DNA-Binding Proteins; D014157:Transcription Factors; D025721:WT1 Proteins