The Cut-Homeodomain Transcriptional Activator HNF-6 Is Coexpressed with Its Target Gene HNF-3β in the Developing Murine Liver and Pancreas

Abstract

AbstractMurine hepatocyte nuclear factor-3β (HNF-3β) protein is a member of a large family of developmentally regulated transcription factors that share homology in the winged helix/fork headDNA binding domain and that participate in embryonic pattern formation. HNF-3β also mediates cell-specific transcription of genes important for the function of hepatocytes, intestinal and bronchiolar epithelial, and pancreatic acinar cells. We have previously identified a liver-enriched transcription factor, HNF-6, which is required for HNF-3β promoter activity and also recognizes the regulatory region of numerous hepatocyte-specific genes. In this study we used the yeast one-hybrid system to isolate the HNF-6 cDNA, which encodes a cut-homeodomain-containing transcription factor that binds with the same specificity as the liver HNF-6 protein. Cotransfection assays demonstrate that HNF-6 activates expression of a reporter gene driven by the HNF-6 binding site from either the HNF-3β or transthyretin (TTR) promoter regions. We used interspecific backcross analysis to determine that murineHnf6gene is located in the middle of mouse chromosome 9.In situhybridization studies of staged specific embryos demonstrate that HNF-6 and its potential target gene, HNF-3β, are coexpressed in the pancreatic and hepatic diverticulum. More detailed analysis of HNF-6 and HNF-3β's developmental expression patterns provides evidence of colocalization in hepatocytes, intestinal epithelial, and in the pancreatic ductal epithelial and exocrine acinar cells. The expression patterns of these two transcription factors do not overlap in other endoderm-derived tissues or the neurotube. We also found that HNF-6 is also abundantly expressed in the dorsal root ganglia, the marginal layer, and the midbrain. At day 18 of gestation and in the adult pancreas, HNF-6 and HNF-3β transcripts colocalize in the exocrine acinar cells, but their expression patterns diverge in other pancreatic epithelium. HNF-6, but not HNF-3β, expression continues in the pancreatic ductal epithelium, whereas only HNF-3β becomes restricted to the endocrine cells of the islets of Langerhans. We discuss these expression patterns with respect to specification of hepatocytes and differentiation of the endocrine and exocrine pancreas