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SELEX and missing phosphate contact analyses reveal flexibility within the AP-2[alpha] protein: DNA binding complex
N Mohibullah1, A Donner, J A Ippolito
1Department of Molecular Cellular and Developmental Biology, Yale University, 266 Whitney Avenue, New Haven, CT 06511, USA.
Nucleic Acids Research
|June 22, 1999
Summary
The AP-2 transcription factor family
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- AP-2 transcription factors are crucial for vertebrate development and implicated in cancer.
- The specific DNA sequences recognized by AP-2 proteins are not well-defined.
- Understanding AP-2 binding sites is key to identifying its target genes.
Purpose of the Study:
- To identify novel AP-2 binding sites using SELEX analysis.
- To characterize the AP-2 protein:DNA interactions and binding site requirements.
- To refine the consensus sequences recognized by AP-2[alpha].
Main Methods:
- Systematic Evolution of Ligands by Exponential Enrichment (SELEX) was employed to discover new AP-2 binding sites.
- Missing phosphate and nucleotide competition assays were used to map protein:DNA contacts.
- Sequence analysis of identified binding sites to determine consensus motifs.
Main Results:
- Multiple new AP-2[alpha] binding sites were identified.
- The AP-2[alpha] protein:DNA complex exhibits flexibility.
- AP-2[alpha] recognizes three related sequence motifs: GCC N3 GGC, GCC N4 GGC, and GCC N3/4 GGG.
Conclusions:
- Refined consensus sequences for AP-2[alpha] binding have been established.
- These findings will aid in identifying novel target genes regulated by AP-2[alpha].
- This research advances the understanding of AP-2 mediated gene regulation.