A novel element regulates expression of the proximal human proglucagon promoter in islet cells

The human and rat proglucagon gene proximal promoter regions have differing transcriptional activities in pancreatic islet cell lines, with 300 bases of rat proglucagon 5′ flanking sequence being sufficient to support expression in rodent islet cell lines, while the homologous human sequences are transcriptionally silent. To better understand the changes in promoter activity between human and rat we have used a comparative approach and cloned promoters from diverse mammalian species and tested their transcriptional activities. Proglucagon gene proximal promoter regions from species representing three orders of mammals (rodents, artiodactyls, and carnivores) support transcription in rodent islet cell lines, while promoters from primates (human and rhesus monkey), despite significant sequence conservation, failed to drive reporter gene expression. These results suggest that nucleotide changes have occurred to the sequence of the proximal promoter region of the proglucagon gene during the evolution of primates that prevent them from supporting expression in rodent islet cell lines. Using hybrid human-rat proglucagon promoters and site-directed mutagenesis we identified a novel regulatory element in the human proglucagon proximal promoter, located between the G2 and G3 enhancer elements that is responsible for most of the difference in transcriptional activity between the human and rat proximal proglucagon promoters.