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We demonstrate that a factor (PO-B), detected in a number of mammalian cell lines, binds specifically between the TATA box and the transcription initiation site of the pituitary-specific pro-opiomelanocortin (POMC) gene. Methylation interference and DNAse 1 footprint analysis revealed that the binding site of this protein, -3 to -15, does not overlap the POMC TATA box or cap site. Mutation of the PO-B binding site significantly decreased the transcriptional activity of the POMC promoter after transient transfection into the ATt-20 mouse pituitary tumor cell line and also in in vitro transcription assays. In contrast, mutation of the TATA box produced no overall decrease in transcription in vivo but induced multiple heterogeneous start sites in vitro. A vector harboring the PO-B site alone was unable to direct transcription initiation. PO-B represents a new transcription factor that may be able to facilitate POMC gene expression by interacting with components of the transcription initiation complex.

A novel transcriptional activator (PO-B) binds between the TATA box and cap site of the pro-opiomelanocortin gene