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Identification of a novel AP-2 consensus DNA binding site
M S Gee1, C J Sarkisian, W S el-Deiry
1Howard Hughes Medical Institute, Department of Medicine and Genetics, University of Pennsylvania School of Medicine, Philadelphia 19104, USA.
Biochemical and Biophysical Research Communications
|February 25, 1998
Summary
Researchers identified novel DNA binding sites for Activator Protein (AP)-2, a crucial transcription factor for mouse development. This discovery refines our understanding of AP-2
Area of Science:
- Molecular Biology
- Genetics
- Cancer Research
Background:
- Activator Protein (AP)-2 is a vital transcription factor essential for mammalian development.
- AP-2 regulates genes involved in cell growth, including erbB-2 and p21 WAF1/CIP1.
- The p21 WAF1/CIP1 gene's induction is linked to inhibiting human cancer cell proliferation.
Purpose of the Study:
- To precisely define the DNA-protein interactions of AP-2.
- To identify novel AP-2 binding sites beyond known consensus sequences.
Main Methods:
- Cyclic Amplification and Selection of Targets (CAST'ing) was employed using random oligonucleotides and recombinant human AP-2.
- Electrophoretic mobility shift assays (EMSAs) were used to validate AP-2 binding to identified sites.
Main Results:
- Seventeen novel AP-2 binding sites were identified.
- High-affinity and specific AP-2 binding was observed at several novel sites, distinct from previously known sequences.
- A new putative consensus binding sequence (5'-TAGAAAGNYCYNG-3') was proposed based on novel high-affinity sites.
Conclusions:
- The identified novel DNA binding sites expand the known targets of AP-2.
- This research provides a more precise understanding of AP-2's DNA binding specificity.
- The findings may facilitate the discovery of new AP-2-regulated genes and pathways.