Role of signal transducers and activators of transcription 1 and -3 in inducible regulation of the human angiotensinogen gene by interleukin-6

C. T. Sherman, A. R. Brasier

Research output: Contribution to journalArticle

44 Citations (Scopus)

Abstract

The circulating level of angiotensinogen (AGT) is dynamically regulated as an important determinant of blood pressure and electrolyte homeostasis. Because the mechanisms controlling the regulated expression of human angiotensinogen (hAGT) are unknown, we investigated the inducible regulation of the hAGT gene in well differentiated HepG2 cells. Interleukin-6 (IL-6) stimulation produced a 3.2-fold increase in hAGT mRNA peaking at 96 h after stimulation. Deletional mutagenesis of the hAGT promoter in transient transfection assays identified an IL-6 response domain between nucleotides -350 and -122 containing three reiterated motifs, termed human acute phase response elements (hAPREs). Although mutation of each site individually caused a fall in IL-6-inducible luciferase activity, mutation of all three sites was required to block the IL-6 effect. Electrophoretic mobility shift assay (EMSA), supershift, and microaffinity DNA binding assays indicate IL-6-inducible high-affinity binding of signal transducers and activators of transcription 1 and -3 (STAT1 and -3) to hAPRE1 and -3 but only low-affinity binding to hAPRE2. Expression of a dominant-negative form of STAT3, but not STAT1, produced a concentration-dependent reduction in IL-6-induced hAGT transcription and endogenous mRNA expression. These data indicate that STAT3 plays a major role in hAGT gene induction through three functionally distinct hAPREs in its promoter and suggest a mechanism for its up-regulation during the acute-phase response.

Original languageEnglish (US)
Pages (from-to)441-457
Number of pages17
JournalMolecular Endocrinology
Volume15
Issue number3
DOIs
StatePublished - 2001

Fingerprint

STAT1 Transcription Factor
STAT3 Transcription Factor
Angiotensinogen
Interleukin-6
Acute-Phase Reaction
Genes
Response Elements
Messenger RNA
Mutation
Hep G2 Cells
Electrophoretic Mobility Shift Assay
Luciferases
Mutagenesis
Electrolytes
Transfection
Homeostasis
Up-Regulation
Nucleotides
Blood Pressure

ASJC Scopus subject areas

  • Molecular Biology
  • Endocrinology, Diabetes and Metabolism

Cite this

Role of signal transducers and activators of transcription 1 and -3 in inducible regulation of the human angiotensinogen gene by interleukin-6. / Sherman, C. T.; Brasier, A. R.

In: Molecular Endocrinology, Vol. 15, No. 3, 2001, p. 441-457.

Research output: Contribution to journalArticle

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