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Related Experiment Videos

Thyroid Hormone Stimulates Renin Gene Expression Through the Thyroid Hormone Response Element.

Hiroyuki Kobori1, Matsuhiko Hayashi, Takao Saruta

  • 1Department of Internal Medicine, Keio University School of Medicine, Tokyo, Japan.

Hypertension (Dallas, Tex. : 1979)
|February 24, 2001
PubMed
Summary

Thyroid hormone stimulates human renin gene promoter activity via a specific DNA sequence. This finding reveals a thyroid hormone response element crucial for regulating renin gene expression in cells.

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Area of Science:

  • Endocrinology
  • Molecular Biology
  • Genetics

Background:

  • Thyroid hormone is known to influence renin synthesis.
  • The precise mechanisms by which thyroid hormone regulates the human renin gene are not fully understood.

Purpose of the Study:

  • To investigate the 5'-flanking sequence of the human renin gene for thyroid hormone-responsive promoter activity.
  • To identify potential thyroid hormone response elements (TREs) within the human renin gene promoter.

Main Methods:

  • Utilized Calu-6 cells, which express thyroid hormone receptor-beta and secrete renin.
  • Employed luciferase reporter gene assays with various lengths of the human renin gene's 5'-flanking sequence.
  • Performed electrophoretic mobility shift assays (EMSAs) to assess protein binding to DNA fragments.

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Main Results:

  • Thyroid hormone (3,3',5-Triiodo-L-thyronine) significantly stimulated promoter activity for constructs containing sequences between -1111/+12 and -1298/+12 of the human renin gene.
  • No significant stimulation was observed with shorter promoter constructs.
  • A specific DNA sequence (5'-AGG TCA GGT CAc aat GTT CCT-3') between nucleotides -1111 and -953 was identified as a potential TRE.
  • Site-directed mutations within this sequence abolished thyroid hormone-mediated promoter activation and altered basal promoter activity.
  • EMSAs confirmed that nuclear proteins from Calu-6 cells bind to the intact -1111/-953 DNA fragment, but not to mutated versions.

Conclusions:

  • Thyroid hormone stimulates the promoter activity of the human renin gene in Calu-6 cells.
  • This stimulation is mediated through a thyroid hormone response element located between nucleotides -1111 and -953 of the human renin gene.
  • The identified TRE is essential for both basal promoter activity and thyroid hormone-induced activation, involving binding of nuclear proteins.