A set of polyclonal and monoclonal antibodies reveals major differences in post-translational modification of the rat HNF1 and vHNF1 homeoproteins

Abstract The related homeodomain-containing transcription factors HNF1 (HNF1α) and vHNF1 (HNF1β) recognise common target DNA sequences in the regulatory regions of many genes and are expressed in several parenchymal cell types, predominantly in liver, kidney, intestine and pancreas. HNFI-mull mutant mice, with a wild-type vHNF1 gene, develop normally, but die within a few weeks of birth with severe liver and kidney failure. Humans with a mutation in the HNF1α gene develop non-insulin dependent diabetes on maturity (MODY 3). To determine distinctive roles for each of these proteins we produced a set of polyclonal sera and monoclonal antibodies, directed against different parts of the rat HNF1 and vHNF1 proteins. These antibodies reveal that HNF1 is present in vivo as a heterogeneous mixture of 92–98 kDa molecular mass polypeptides, a mass higher than that expected from its amino acid sequence, vHNF1 is present in the form of two isoforms of roughly the expected molecular masses, 65 and 68 kDa. In addition, some antibodies prepared against bacterially produced HNF1 recognise vHNF1 but not HNF1, in liver and kidney extracts. Hence, we present the first evidence for differential post-translational modification of HNF1 and vHNF1 proteins.