Insulin upstream factor 1 and a novel ubiquitous factor bind to the human islet amyloid polypeptide/amylin gene promoter.
The islet amyloid polypeptide (IAPP) gene is expressed primarily in the islet beta-cell and the peptide is co-secreted with insulin. To investigate mechanisms important in its regulation, we have used the electrophoretic mobility-shift assay and methylation interference to determine systematically sites of DNA-protein interactions in the human IAPP promoter. We identified beta-cell-specific DNA-protein complexes at three sites, each of which contained a consensus binding site for insulin upstream factor I (IUF-I). This complex was displaced with an antiserum to IUF-1, confirming that IUF-1 binds to the human IAPP promoter in vitro. We have also identified a DNA-protein complex within the region -220/-250 in both beta- and non-beta-cell lines. This region contains a motif with partial identity with the binding site for the ubiquitous transcription factor upstream stimulatory factor (USF), which binds to the human insulin promoter. However, purified USF was not able to bind to this putative site in the IAPP promoter and an oligonucleotide containing a functional USF-binding site was unable to displace binding from the IAPP oligonucleotide. Methylation interference revealed that the DNA-protein complex binds to a sequence that overlaps the USE-like sequence, and may therefore be a novel helix-loop-helix protein. These results suggest that, although both IAPP and insulin are beta-cell peptides, IAPP contains regulatory regions both common to and distinct from insulin.
Bretherton-Watt
D
D
Division of Biochemistry, School of Biological Sciences, University of Manchester, U.K.
Gore
N
N
Boam
D S
DS
D000682:Amyloid / Q000235:genetics*; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002478:Cells, Cultured; D004268:DNA-Binding Proteins / Q000378:metabolism; D006801:Humans; D007328:Insulin / Q000378:metabolism*; D058228:Islet Amyloid Polypeptide; D007515:Islets of Langerhans / Q000378:metabolism; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D011401:Promoter Regions, Genetic; D051381:Rats
D000682:Amyloid; D004268:DNA-Binding Proteins; D007328:Insulin; D058228:Islet Amyloid Polypeptide; D009838:Oligodeoxyribonucleotides