biochemical characterization of diverse stat5b-binding enhancers that mediate growth hormone-activated insulin-like growth factor-i gene transcription生化特征的多样化stat5b-binding增强剂,调解增长hormone-activated胰岛素样生长因子基因转录.pdfVIP

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biochemical characterization of diverse stat5b-binding enhancers that mediate growth hormone-activated insulin-like growth factor-i gene transcription生化特征的多样化stat5b-binding增强剂,调解增长hormone-activated胰岛素样生长因子基因转录.pdf

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biochemical characterization of diverse stat5b-binding enhancers that mediate growth hormone-activated insulin-like growth factor-i gene transcription生化特征的多样化stat5b-binding增强剂,调解增长hormone-activated胰岛素样生长因子基因转录

Biochemical Characterization of Diverse Stat5b-Binding Enhancers That Mediate Growth Hormone-Activated Insulin-Like Growth Factor-I Gene Transcription 1 1 1 2 1 Ben Varco-Merth , Kasim Mirza , Damir T. Alzhanov , Dennis J. Chia , Peter Rotwein * 1 Department of Biochemistry and Molecular Biology, Oregon Health Science University, Portland, Oregon, United States of America, 2 Department of Pediatrics, Mt. Sinai School of Medicine, New York, New York, United States of America Abstract Many of the biological effects of growth hormone (GH) are mediated by insulin-like growth factor I (IGF-I), a 70-amino acid secreted peptide whose gene expression is rapidly induced by GH via the Stat5b transcription factor. We previously identified multiple evolutionarily conserved GH-activated chromosomal binding domains for Stat5b within the rat Igf1 locus, and proposed that they could regulate IGF-I gene activity. Here we investigate the biochemical and functional characteristics of these putative long-range transcriptional enhancers. Each element contained 2 or 3 individual Stat5b recognition sequences that could bind Stat5b in vitro, but with affinities that varied over a .100-fold range. Full transcriptional responsiveness to GH required that all Stat5b sites be intact within an individual enhancer. Replacement of a single lower-affinity Stat5b sequence with a higher-affinity one increased in vitro binding of Stat5b, and boosted transcriptional potency of the entire element to GH. As enhanced transcriptional activity involved changes in only one or two nucleotides within an enhancer DNA segment, there appears to be remarkable specificity and