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Related Experiment Videos

The rat growth hormone gene contains multiple thyroid response elements.

M F Norman1, T N Lavin, J D Baxter

  • 1Metabolic Research Unit, University of California, San Francisco 94143.

The Journal of Biological Chemistry
|July 15, 1989
PubMed
Summary
This summary is machine-generated.

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Researchers identified four high-affinity binding sites for the thyroid hormone receptor on rat growth hormone DNA. These sites are crucial for regulating gene expression in response to thyroid hormone.

Area of Science:

  • Molecular Biology
  • Genetics
  • Endocrinology

Background:

  • Thyroid hormone receptor (TR) regulates gene transcription.
  • Understanding TR binding to target genes is key to its function.

Purpose of the Study:

  • To identify and characterize TR binding sites in the rat growth hormone (GH) gene promoter.
  • To investigate the role of these binding sites in thyroid hormone-mediated gene regulation.

Main Methods:

  • Gel retardation assays using purified rat liver TR.
  • Methylation interference and methidium-propyl-EDTA (MPE) footprinting.
  • Site-directed mutagenesis of identified binding sites.
  • Reporter gene assays (chloramphenicol acetyltransferase) in pituitary cells.

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Main Results:

  • Four high-affinity TR binding sites were mapped in the rat GH gene 5'-flanking DNA (-180, -160, -60, -20).
  • A conserved purine-rich sequence (GGGATCGC) was identified within these sites.
  • Mutations in upstream sites decreased TR binding affinity.
  • Mutations at all four sites reduced thyroid hormone responsiveness of the GH promoter.

Conclusions:

  • Multiple TR binding elements cooperate to control rat GH gene responsiveness to thyroid hormone.
  • Common DNA sequences within these elements may be critical for TR-DNA recognition.