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Molecular Endocrinology 14 (1):99
Copyright © 2000 by The Endocrine Society

Angiotensin II Induces Nuclear Factor (NF)-{kappa}B1 Isoforms to Bind the Angiotensinogen Gene Acute-Phase Response Element: A Stimulus-Specific Pathway for NF-{kappa}B Activation

Mohammad Jamaluddin, Tao Meng, Juan Sun, Istvan Boldogh, Youqi Han and Allan R. Brasier

Department of Internal Medicine (M.J., T.M., J.S., Y.H., A.R.B.) Department of Microbiology & Immunology (I.B.) Sealy Center for Molecular Sciences (A.R.B.) University of Texas Medical Branch Galveston, Texas 77555-1060

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} (TNF{alpha})]- inducible enhancer by binding the heterodimeric nuclear factor-{kappa}B (NF-{kappa}B) complex Rel A•NF-{kappa}B1. Here, we compare the mechanism for NF-{kappa}B activation by the AII agonist, Sar1 AII, with TNF{alpha} in HepG2 hepatocytes. Although Sar1 AII and TNF{alpha} both rapidly activate APRE-driven transcription within 3 h of treatment, the pattern of inducible NF-{kappa}B binding activity in electrophoretic mobility shift assay is distinct. In contrast to the TNF{alpha} mechanism, which strongly induces Rel A•NF-{kappa}B1 binding, Sar1 AII selectively activates a heterogenous pattern of NF-{kappa}B1 binding. Using a two-step microaffinity DNA binding assay, we observe that Sar1 AII recruits 50-, 56-, and 96-kDa NF-{kappa}B1 isoforms to bind the APRE. Binding of all three NF-{kappa}B1 isoforms occurs independently of changes in their nuclear abundance or proteolysis of cytoplasmic I{kappa}B inhibitors. Phorbol ester-sensitive protein kinase C (PKC) isoforms are required because PKC down-regulation completely blocks AII-inducible transcription and inducible NF-{kappa}B1 binding. We conclude that AII stimulates the NF-{kappa}B transcription factor pathway by activating latent DNA-binding activity of NF-{kappa}B subunits through a phorbol ester-sensitive (PKC-dependent) mechanism.




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