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c-Myc target gene specificity is determined by a post-DNAbinding mechanism
1McArdle Laboratory for Cancer Research, University of Wisconsin Medical School, Madison, WI 53706, USA.
Summary
This study shows that Myc, a transcription factor, binds to the cad promoter. The cad promoter
Area of Science:
- Molecular Biology
- Genetics
- Cell Biology
Background:
- Identifying transcription factor targets in cells is challenging.
- Understanding protein-DNA interactions is crucial for gene regulation.
Purpose of the Study:
- To investigate Myc and USF1 binding to the endogenous cad promoter.
- To determine the functional consequences of transcription factor binding in vivo.
- To elucidate mechanisms of transcription factor target gene specificity.
Main Methods:
- Utilized formaldehyde crosslinking and Gal4 fusion proteins.
- Performed immunoprecipitation with antibodies against Myc and USF1.
- Analyzed DNA binding and promoter activity in intact mammalian cells.
Main Results:
- Both Myc and USF1 bind to the endogenous cad promoter.
- Only Myc's transactivation domain affects cad promoter activity.
- Myc binding to cad fluctuates with cell growth.
- Myc binding site location dictates functional outcomes.
Conclusions:
- The cad gene is a direct target of Myc.
- Post-DNA-binding mechanisms determine Myc target gene specificity.
- Developed a method to study transcription factor binding to single-copy mammalian genes in vivo.