Identification of a Novel AP-2 Consensus DNA Binding Site

• Published in: Biochemical and biophysical research communications Vol. 243; no. 1; pp. 307 - 316
• Main Authors: Gee, Michael S., Sarkisian, Christopher J., El-Deiry, Wafik S.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 04-02-1998
• Subjects: Animals; Base Sequence; Binding Sites - genetics; Consensus Sequence; DNA - genetics; DNA - metabolism; DNA-Binding Proteins - metabolism; Humans; Mice; Protein Binding; Recombinant Fusion Proteins - metabolism; Transcription Factor AP-2; Transcription Factors - metabolism
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1006/bbrc.1997.8035

Activator Protein (AP)-2 is a transcription factor that is required for mouse development. AP-2 activates expression of positive and negative growth regulators including erbB-2 and p21WAF1/CIP1. Induction of p21 has been correlated with cell cycle and growth inhibition of human cancer cells. Because several endogenous AP-2 binding sites do not fit the known consensus sequences well, we sought to define AP-2's interaction with DNA more precisely. UsingCyclicAmplification andSelection ofTargets (CAST'ing) of random oligonucleotide sequences and recombinant human AP-2 protein, we identified 17 novel AP-2 binding sites. Mobility shift assays showed significant AP-2 binding of the novel sites as compared to p21, erbB-2 and hMtIIa sites. Several sites that bound with high specificity and affinity did not fit known AP-2 consensus sequences. A sequence comparison based on several of the novel sequences yielded a putative consensus binding sequence of 5′-TAGAAAGNYCYNG-3′. These DNA binding sites may help identify novel targets of AP-2 and aid in further understanding AP-2 function.