A novel growth hormone response element unrelated to STAT (signal transducer and activator of transcription)-binding sites is a bifunctional enhancer.

Regulation of gene expression by GH has so far been shown to be mediated by a few cis-acting elements, most of which are signal transducer and activator of transcription (STAT)-binding sites. Here we have characterized a novel GH-response element present in the promoter of rat serine protease inhibitor (spi) genes. It consists of a 13 nucleotide-long GAGA box containing two GAGGAG repeats separated by a G, structurally unrelated to STAT-binding sites. In hepatocytes, the spi GAGA box behaves as a position-dependent bifunctional enhancer controlling basal and GH-dependent transcription. In addition, spi GAGA box oligonucleotides inhibit cell-free transcription driven by GAGA box-containing as well as GAGA box-less promoters, suggesting that the spi GAGA box interacts directly or indirectly with component(s) of the basic transcriptional machinery. Mobility shift assays showed that this GAGA box is specifically recognized by nuclear factors that are unrelated to previously characterized proteins binding to purine-rich elements or to GH-activated STATs. Finally, experiments performed with cells expressing wild type, truncated, or mutated forms of the GH receptor indicate that protein kinase Janus kinase 2 is involved in the GH-dependent activation of the spi GAGA box. These studies reveal the existence of an as yet unidentified Janus kinase-2-dependent, STAT-independent pathway in GH activation of gene expression.