Involvement of a novel Rac/RhoA guanosine triphosphatase-nuclear factor-κB inducing kinase signaling pathway mediating angiotensin II-induced RelA transactivation

Sanjeev Choudhary, Muping Lu, Ruwen Cui, Allan R. Brasier

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Angiotensin II (Ang II) is the major effector peptide of the renin angiotensin system that induces inflammatory gene expression through the nuclear factor-κB (NF-κB) transcription factor. Activation of latent cytoplasmic NF-κB is controlled by distinct pathways, the best known being the canonical pathway controlling IκB kinase activation. Interestingly, Ang II only weakly activates the canonical pathway. Although basal nucleocytoplasmic RelA shuttling is required for Ang II stimulation, changes in RelA translocation do not account for its transcriptional effect. Instead, Ang II rapidly induced RelA phosphorylation at Ser residue 536, and complex formation with the Ser536 kinase known as the NF-κB-inducing kinase (NIK)/MEKK14. The requirement of NIK in Ang II-inducible transcription was shown by expressing a dominant-negative NIK or small interfering RNA (siRNA)-mediated knockdown; both inhibited Ang II-induced transcription. Conversely, constitutively active NIK potently induced RelA transactivation activity. Consistent with its actions independent of the canonical pathway, NIK induces the activity of the RelA transactivation domains -1 and -2 in constitutively nuclear GAL4-RelA fusion proteins that do not bind IκBα. Ang II induces NIK activity, phosphorylation of its endogenous IκB kinase α substrate, and induction of nuclear NF-κB2 (p52) processing. NIK down-regulation prevents Ang II-induced phospho-Ser 536 RelA formation, indicating that it is essential for RelA activation. The Ang II pathway further involves the RhoA small GTP-binding protein because RhoA inhibition blocks Ang II-induced transcriptional activity and formation of phospho-Ser536 RelA formation. Finally, we demonstrate that Ang II infusion in vivo rapidly induces phospho-Ser 536 RelA formation and activation of the NF-κB-dependent IL-6 gene. These data indicate that Ang II induces NF-κB-dependent transcription through an alternative pathway, being largely independent of IκB proteolysis, but mediated by the small GTPases Rac/RhoA, required for NIK·RelA complex formation and inducible Ser536 RelA phosphorylation.

Original languageEnglish (US)
Pages (from-to)2203-2217
Number of pages15
JournalMolecular Endocrinology
Volume21
Issue number9
DOIs
StatePublished - Sep 2007

Fingerprint

Guanosine
Angiotensin II
Transcriptional Activation
Phosphotransferases
Phosphorylation
rhoA GTP-Binding Protein
Monomeric GTP-Binding Proteins
Renin-Angiotensin System
Small Interfering RNA
Proteolysis
Interleukin-6
Transcription Factors
Down-Regulation

ASJC Scopus subject areas

  • Molecular Biology
  • Endocrinology, Diabetes and Metabolism

Cite this

Involvement of a novel Rac/RhoA guanosine triphosphatase-nuclear factor-κB inducing kinase signaling pathway mediating angiotensin II-induced RelA transactivation. / Choudhary, Sanjeev; Lu, Muping; Cui, Ruwen; Brasier, Allan R.

In: Molecular Endocrinology, Vol. 21, No. 9, 09.2007, p. 2203-2217.

Research output: Contribution to journalArticle

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abstract = "Angiotensin II (Ang II) is the major effector peptide of the renin angiotensin system that induces inflammatory gene expression through the nuclear factor-κB (NF-κB) transcription factor. Activation of latent cytoplasmic NF-κB is controlled by distinct pathways, the best known being the canonical pathway controlling IκB kinase activation. Interestingly, Ang II only weakly activates the canonical pathway. Although basal nucleocytoplasmic RelA shuttling is required for Ang II stimulation, changes in RelA translocation do not account for its transcriptional effect. Instead, Ang II rapidly induced RelA phosphorylation at Ser residue 536, and complex formation with the Ser536 kinase known as the NF-κB-inducing kinase (NIK)/MEKK14. The requirement of NIK in Ang II-inducible transcription was shown by expressing a dominant-negative NIK or small interfering RNA (siRNA)-mediated knockdown; both inhibited Ang II-induced transcription. Conversely, constitutively active NIK potently induced RelA transactivation activity. Consistent with its actions independent of the canonical pathway, NIK induces the activity of the RelA transactivation domains -1 and -2 in constitutively nuclear GAL4-RelA fusion proteins that do not bind IκBα. Ang II induces NIK activity, phosphorylation of its endogenous IκB kinase α substrate, and induction of nuclear NF-κB2 (p52) processing. NIK down-regulation prevents Ang II-induced phospho-Ser 536 RelA formation, indicating that it is essential for RelA activation. The Ang II pathway further involves the RhoA small GTP-binding protein because RhoA inhibition blocks Ang II-induced transcriptional activity and formation of phospho-Ser536 RelA formation. Finally, we demonstrate that Ang II infusion in vivo rapidly induces phospho-Ser 536 RelA formation and activation of the NF-κB-dependent IL-6 gene. These data indicate that Ang II induces NF-κB-dependent transcription through an alternative pathway, being largely independent of IκB proteolysis, but mediated by the small GTPases Rac/RhoA, required for NIK·RelA complex formation and inducible Ser536 RelA phosphorylation.",
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