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Antisense transcripts and protein binding motifs within the Wilms tumour (WT1) locus
C E Campbell1, A Huang, A L Gurney
1Department of Cancer Biology, Cleveland Clinic Foundation, Ohio 44195.
Oncogene
|February 1, 1994
Summary
The WT1 gene locus produces two transcripts, WT1 and WIT1, with WT1 acting as a transcriptional regulator. A bidirectional promoter controls their expression, and WT1 protein binds to this regulatory region.
Area of Science:
- Molecular Biology
- Genetics
- Developmental Biology
Background:
- The WT1 locus in mammals is transcriptionally active in specific kidney and gonad epithelial cells.
- WT1 encodes a zinc finger protein, a likely transcriptional regulator.
- A second transcript, WIT1, has an unknown function and undergoes alternative splicing.
Purpose of the Study:
- To investigate the transcriptional regulation of the WT1 locus.
- To characterize the promoter region and its interaction with WT1 protein.
Main Methods:
- Analysis of gene transcripts and alternative splicing.
- Identification of transcriptional start sites.
- Transient transfection assays using CAT gene constructs.
- In vitro binding assays with nuclear extracts and recombinant WT1 protein.
Main Results:
- The WT1 locus exhibits multiple transcriptional start sites for both WT1 and WIT1.
- Some WIT1 transcripts originate from the antisense strand of WT1's first exon.
- A bidirectional promoter region was identified surrounding the transcriptional start sites.
- WT1 protein binds to sequences within this bidirectional promoter region.
Conclusions:
- The WT1 locus is regulated by a bidirectional promoter.
- WT1 protein autoregulates its own transcription by binding to this promoter.
- Complex transcriptional regulation governs the expression of WT1 and WIT1.