Dispersed chromosomal Stat5b-binding elements mediate growth hormone-activated insulin-like growth factor-I gene transcription

Dennis J. Chia, Ben Varco-Merth, Peter Rotwein

Research output: Contribution to journalArticle

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Abstract

The growth hormone (GH)-insulin-like growth factor-I (IGF-I) axis regulates somatic growth during childhood and orchestrates tissue repair throughout the life span. Recently described inactivating mutations in Stat5b in humans with impaired growth have focused attention on this transcription factor as a key agent linking GH-stimulated signals to IGF-I gene expression, and several putative Stat5b sites have been identified in the IGF-I gene. Here, we define and characterize potential GH- and Stat5b-activated chromosomal enhancers that can regulate IGF-I gene transcription. Of 89 recognizable Stat5 sequences in 200 kb centering on the rat IGF-I gene, 22 resided within conserved regions and/or were identical among different species. Only 15 of these sites, organized into 7 distinct domains, were found to bind Stat5b by quantitative chromatin immunoprecipitation assays in liver chromatin of rats, but only after acute GH treatment. These sites could bind Stat5b in vitro, and individual domains could mediate GH- and Stat5b-stimulated IGF-I promoter activity in cultured cells. Further analyses revealed that four Stat5b domains possessed chromatin signatures of enhancers, including binding of co-activators p300 and Med1, and RNA polymerase II. These modifications preceded GH-stimulated recruitment of Stat5b, as did lysine 4 monomethylation of histone H3, which was enriched in 6/7 Stat5bbinding elements. In contrast, histone acetylation was induced by GH but was limited to Stat5b binding domains found within the IGF-I transcription unit. We conclude that GH stimulates recruitment of Stat5b to multiple dispersed regions within the igf1 locus, including several with properties consistent with long range transcriptional enhancers that collectively regulate GH-activated IGF-I gene transcription.

Original languageEnglish (US)
Pages (from-to)17636-17647
Number of pages12
JournalJournal of Biological Chemistry
Volume285
Issue number23
DOIs
StatePublished - Jun 4 2010
Externally publishedYes

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Transcription
Insulin-Like Growth Factor I
Growth Hormone
Genes
Chromatin
Histones
Rats
Acetylation
RNA Polymerase II
Chromatin Immunoprecipitation
Growth
Gene expression
Liver
Lysine
Cultured Cells
Assays
Repair
Transcription Factors
Cells
Tissue

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Dispersed chromosomal Stat5b-binding elements mediate growth hormone-activated insulin-like growth factor-I gene transcription. / Chia, Dennis J.; Varco-Merth, Ben; Rotwein, Peter.

In: Journal of Biological Chemistry, Vol. 285, No. 23, 04.06.2010, p. 17636-17647.

Research output: Contribution to journalArticle

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