Ras controls tumor necrosis factor receptor-associated factor (TRAF)6-dependent induction of nuclear factor-kappa b. Selective regulation through receptor signaling components

J Biol Chem. 2001 Mar 2;276(9):6280-8. doi: 10.1074/jbc.M006772200. Epub 2000 Nov 15.

Abstract

In the present study, we show that Ras activity differentially controls interleukin (IL)-1 induced transcription factor activation by selective regulation of responses mediated by receptor complex components. Initial experiments revealed that stimulation with IL-1 caused a rapid, matrix-dependent activation of Ras. The effect was transient, peaking at 5 min and returning to base levels after 30 min. Activation correlated with pronounced changes in cell shape in EGFPH-Ras transfected cells. Transfection with the dominant negative mutant, Ras(Asn-17), inhibited IL-1 induced activation of the IL-8 promoter as well as of NF-kappa B and AP-1 synthetic promoters in transient transfection assays. Furthermore, overexpression of the IL-1 signaling proteins TRAF6 or MyD88 gave characteristic activation of IL-8, which was accentuated in the presence of IL-1. Co-transfection with Ras(Asn-17) gave a dose-dependent inhibition of TRAF6-induced responses in the presence and absence of IL-1, but had no effect on MyD88 mediated activity. Similarly, induction of NF-kappa B was abolished by Ras(Asn-17) only in TRAF6-transfected cells. In contrast, inhibiting Ras activity limited AP-1-mediated responses through both receptor complex proteins. Constitutively active Ras(Val-12) increased the TRAF6 induced activity of the NF-kappa B pathway similar to the effect induced by IL-1, while the Ras(Val-12) induced activity was not inhibited by co-transfection with a dominant negative TRAF6. Our data show that activation of the Ras GTPase is an early, matrix-dependent response in IL-1 signaling which participates in structural regulation of IL-1-induced genes. In addition, they show that the Ras induced effect selectively regulates TRAF6-mediated activation of the NF-kappa B pathway, suggesting that Ras GTPase represents a convergence point in structural and cytokine responses, with distinct effects on a subset of downstream signaling events.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Adhesion
  • Cells, Cultured
  • Humans
  • Interleukin-1 / pharmacology*
  • Interleukin-8 / genetics
  • Monomeric GTP-Binding Proteins / physiology*
  • NF-kappa B / metabolism*
  • Promoter Regions, Genetic
  • Proteins / physiology*
  • TNF Receptor-Associated Factor 6
  • Transcription, Genetic
  • Transfection

Substances

  • Interleukin-1
  • Interleukin-8
  • NF-kappa B
  • Proteins
  • TNF Receptor-Associated Factor 6
  • Monomeric GTP-Binding Proteins