RGD Reference Report - Insulin-like growth factor I increases rat peptide YY promoter activity through Sp1 binding sites. - Rat Genome Database

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Insulin-like growth factor I increases rat peptide YY promoter activity through Sp1 binding sites.

Authors: Wang, G  Leiter, AB  Englander, EW  Greeley GH, JR 
Citation: Wang G, etal., Endocrinology 2004 Feb;145(2):659-66. Epub 2003 Oct 30.
RGD ID: 1300516
Pubmed: PMID:14592960   (View Abstract at PubMed)
DOI: DOI:10.1210/en.2003-0770   (Journal Full-text)

Studies in rodents demonstrate that the mitogen, IGF-I, stimulates intestinal peptide YY (PYY) expression. To investigate whether the stimulatory influence of IGF-I is exerted at the level of gene transcription, rat PYY 5'-upstream sequences (-2800/+37 bp, -770/+37 bp, -127/+37 bp) fused to the firefly luciferase (luc) reporter gene were transfected into rat pheochromocytoma cells (PC12) and luc activity measured after IGF-I treatment. IGF-I increased transcriptional activity of all constructs similarly; the PYY (-127/+37 bp)-luc construct was used in subsequent experiments. IGF-I increased PYY (-127/+37 bp)-luc activity in a time- and dose-dependent fashion. Sequence analysis detected five putative Sp1 binding sites in the -127/+37-bp sequence. EMSA and supershift experiments using two oligonucleotide fragments of the -127/+37 region showed that Sp1 and Sp3 proteins bound to putative Sp1 sites. Overexpression of Sp1 greatly increased PYY (-127/+37 bp)-luc activity and site-directed mutagenesis of putative Sp1 binding sites decreased basal and IGF-I-induced elevations in PYY (-127/+37 bp)-luc activity. IGF-I treatment also increased Sp1 protein levels and binding activity. Blockade of the IGF-I receptor (IGF-IR) with an IGF-IR antibody decreased the stimulatory influence of IGF-I on Sp1 protein levels and PYY (-127/+37 bp)-luc activity. Together, these findings indicate that IGF-I functions as a positive regulator of PYY gene expression and that the stimulatory effect may be mediated by Sp1 proteins that bind to the proximal PYY promoter region.

Objects referenced in this article
Gene Igf1 insulin-like growth factor 1 Rattus norvegicus
Gene Pyy peptide YY Rattus norvegicus
Gene Sp1 Sp1 transcription factor Rattus norvegicus

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