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Identification of an antisense WT1 promoter in intron 1: implications for WT1 gene regulation
K T Malik1, J I Wallace, S M Ivins
1Department of Pathology and Microbiology, School of Medical Sciences, University of Bristol, UK.
Abstract:
Expression of the Wilms' tumour suppressor gene, WT1, is tightly regulated during nephrogenesis, and loss of function of its gene product correlates with malignancy. By using luciferase reporter gene constructs containing DNA sequences from the first intron of the WT1 gene, we have identified an antisense WT1 promoter operational in the opposite direction relative to the 5' promoter. Transcription directed by the promoter is negatively regulated by upstream elements, but is activated by expression of WT1. This effect of WT1 is reciprocal to that observed on the 5' promoter, suggesting that antisense promoter activity is involved in WT1 gene regulation. By mimicking expression of the transcript regulated by the antisense promoter, we demonstrate that cellular levels of WT1 can be effectively down-regulated by antisense mRNA complementary to sequences in the first exon of WT1.
Insights
Researchers discovered a new WT1 gene promoter that regulates Wilms' tumour suppressor gene expression. This antisense promoter activity can be used to down-regulate WT1 levels, offering potential therapeutic strategies.
Area of Science:
- Molecular Biology
- Genetics
- Cancer Research
Background:
- The Wilms' tumour suppressor gene (WT1) is crucial for kidney development and its proper regulation is vital.
- Dysregulation or loss of WT1 function is linked to Wilms' tumour, a pediatric kidney cancer.
Purpose of the Study:
- To investigate novel regulatory mechanisms of the WT1 gene.
- To identify and characterize a potential antisense promoter for WT1.
Main Methods:
- Utilized luciferase reporter gene constructs with DNA sequences from the WT1 gene's first intron.
- Analyzed transcriptional activity and regulatory elements of the identified antisense promoter.
- Mimicked antisense transcript expression to assess WT1 down-regulation.
Main Results:
- Identified a functional antisense WT1 promoter operating in the opposite direction to the main 5' promoter.
- Demonstrated that WT1 expression reciprocally regulates its own antisense promoter activity.
- Showed that antisense mRNA targeting WT1's first exon effectively reduces cellular WT1 levels.
Conclusions:
- The WT1 gene possesses a reciprocal antisense regulatory mechanism involving a novel promoter.
- Antisense WT1 promoter activity plays a role in the intricate regulation of WT1 gene expression.
- Targeting WT1 with antisense mRNA offers a potential strategy for down-regulating its expression in relevant contexts.