Angiotensin II induces nuclear factor (NF)-kappaB1 isoforms to bind the angiotensinogen gene acute-phase response element: a stimulus-specific pathway for NF-kappaB activation

Mol Endocrinol. 2000 Jan;14(1):99-113. doi: 10.1210/mend.14.1.0400.

Abstract

The vasopressor angiotensin II (AII) activates transcriptional expression of its precursor, angiotensinogen. This biological "positive feedback loop" occurs through an angiotensin receptor-coupled pathway that activates a multihormone-responsive enhancer of the angiotensinogen promoter, termed the acute-phase response element (APRE). Previously, we showed that the APRE is a cytokine [tumor necrosis factor-alpha (TNFalpha)]- inducible enhancer by binding the heterodimeric nuclear factor-kappaB (NF-kappaB) complex Rel A x NF-kappaB1. Here, we compare the mechanism for NF-kappaB activation by the AII agonist, Sar1 AII, with TNFalpha in HepG2 hepatocytes. Although Sar1 AII and TNFalpha both rapidly activate APRE-driven transcription within 3 h of treatment, the pattern of inducible NF-kappaB binding activity in electrophoretic mobility shift assay is distinct. In contrast to the TNFalpha mechanism, which strongly induces Rel A x NF-kappaB1 binding, Sar1 AII selectively activates a heterogenous pattern of NF-kappaB1 binding. Using a two-step microaffinity DNA binding assay, we observe that Sar1 AII recruits 50-, 56-, and 96-kDa NF-kappaB1 isoforms to bind the APRE. Binding of all three NF-kappaB1 isoforms occurs independently of changes in their nuclear abundance or proteolysis of cytoplasmic IkappaB inhibitors. Phorbol ester-sensitive protein kinase C (PKC) isoforms are required because PKC down-regulation completely blocks AII-inducible transcription and inducible NF-kappaB1 binding. We conclude that AII stimulates the NF-kappaB transcription factor pathway by activating latent DNA-binding activity of NF-kappaB subunits through a phorbol ester-sensitive (PKC-dependent) mechanism.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Angiotensin II / analogs & derivatives
  • Angiotensin II / pharmacology*
  • Angiotensinogen / genetics*
  • Angiotensinogen / metabolism*
  • Base Sequence
  • Dose-Response Relationship, Drug
  • Gene Expression Regulation
  • Humans
  • Kinetics
  • Ligases / metabolism
  • Models, Biological
  • Molecular Sequence Data
  • NF-kappa B / metabolism*
  • Protein Binding
  • Protein Isoforms / metabolism
  • Protein Kinase C / metabolism
  • Response Elements
  • Signal Transduction
  • Tetradecanoylphorbol Acetate / pharmacology
  • Time Factors
  • Transcription, Genetic
  • Transfection
  • Tumor Cells, Cultured
  • Tumor Necrosis Factor-alpha / metabolism

Substances

  • NF-kappa B
  • Protein Isoforms
  • Tumor Necrosis Factor-alpha
  • Angiotensinogen
  • Angiotensin II
  • angiotensin II, Sar(1)-
  • Protein Kinase C
  • Ligases
  • guanosine 3',5'-polyphosphate synthetases
  • Tetradecanoylphorbol Acetate