Heat shock inhibits phosphorylation of I-κBα

Thomas P. Shanley, Marnie A. Ryan, Tonyia Eaves-Pyles, Hector R. Wong

Research output: Contribution to journalArticle

50 Citations (Scopus)

Abstract

Previous studies demonstrated that induction of the heat shock response is associated with inhibition of the proinflammatory transcription factor NF-κB by a mechanism involving inhibition of I-κBα degradation. To provide further insight regarding the interactions of these fundamental cellular responses, the present experiments were designed to elucidate the mechanism(s) by which heat shock inhibits degradation of I-κBα. In an in vitro model of inflammatory cell signaling, treatment of RAW 264.7 murine macrophages with LPS (100 ng/mL) caused rapid degradation of I-κBα. Heat shock, 1 h before treatment with LPS, completely inhibited LPS-mediated degradation of I-κBα. Immunoprecipitation studies demonstrated that heat shock inhibited LPS-mediated ubiquitination of I-κBα. Western-blot analyses using a phosphorylated I-κBα-specific antibody demonstrated that heat shock inhibited LPS-mediated phosphorylation of I-κBα. In contrast, heat shock induced phosphorylation of c-jun. In murine fibroblasts having genetic ablation of the heat shock factor-1 gene, heat shock inhibited tumor necrosis factor-α mediated degradation of I-κBα. We conclude that the mechanism by which heat shock inhibits LPS-mediated degradation of I-κBα involves specific inhibition of I-κBα phosphorylation and subsequent I-κBα ubiquitination. In addition, this mechanism does not involve activation of heat shock factor-1 or the heat shock proteins regulated by heat shock factor-1.

Original languageEnglish (US)
Pages (from-to)447-450
Number of pages4
JournalShock
Volume14
Issue number4
StatePublished - Oct 2000
Externally publishedYes

Fingerprint

Shock
Hot Temperature
Phosphorylation
Ubiquitination
Heat-Shock Response
Heat-Shock Proteins
Immunoprecipitation
Transcription Factors
Tumor Necrosis Factor-alpha
Fibroblasts
Western Blotting
Macrophages
Antibodies
Genes

Keywords

  • Heat shock factor-1
  • Inflammation
  • Lipopolysaccharide
  • NF-κB
  • Signal transduction
  • Stress response

ASJC Scopus subject areas

  • Physiology
  • Critical Care and Intensive Care Medicine

Cite this

Shanley, T. P., Ryan, M. A., Eaves-Pyles, T., & Wong, H. R. (2000). Heat shock inhibits phosphorylation of I-κBα. Shock, 14(4), 447-450.

Heat shock inhibits phosphorylation of I-κBα. / Shanley, Thomas P.; Ryan, Marnie A.; Eaves-Pyles, Tonyia; Wong, Hector R.

In: Shock, Vol. 14, No. 4, 10.2000, p. 447-450.

Research output: Contribution to journalArticle

Shanley, TP, Ryan, MA, Eaves-Pyles, T & Wong, HR 2000, 'Heat shock inhibits phosphorylation of I-κBα', Shock, vol. 14, no. 4, pp. 447-450.
Shanley TP, Ryan MA, Eaves-Pyles T, Wong HR. Heat shock inhibits phosphorylation of I-κBα. Shock. 2000 Oct;14(4):447-450.
Shanley, Thomas P. ; Ryan, Marnie A. ; Eaves-Pyles, Tonyia ; Wong, Hector R. / Heat shock inhibits phosphorylation of I-κBα. In: Shock. 2000 ; Vol. 14, No. 4. pp. 447-450.
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AB - Previous studies demonstrated that induction of the heat shock response is associated with inhibition of the proinflammatory transcription factor NF-κB by a mechanism involving inhibition of I-κBα degradation. To provide further insight regarding the interactions of these fundamental cellular responses, the present experiments were designed to elucidate the mechanism(s) by which heat shock inhibits degradation of I-κBα. In an in vitro model of inflammatory cell signaling, treatment of RAW 264.7 murine macrophages with LPS (100 ng/mL) caused rapid degradation of I-κBα. Heat shock, 1 h before treatment with LPS, completely inhibited LPS-mediated degradation of I-κBα. Immunoprecipitation studies demonstrated that heat shock inhibited LPS-mediated ubiquitination of I-κBα. Western-blot analyses using a phosphorylated I-κBα-specific antibody demonstrated that heat shock inhibited LPS-mediated phosphorylation of I-κBα. In contrast, heat shock induced phosphorylation of c-jun. In murine fibroblasts having genetic ablation of the heat shock factor-1 gene, heat shock inhibited tumor necrosis factor-α mediated degradation of I-κBα. We conclude that the mechanism by which heat shock inhibits LPS-mediated degradation of I-κBα involves specific inhibition of I-κBα phosphorylation and subsequent I-κBα ubiquitination. In addition, this mechanism does not involve activation of heat shock factor-1 or the heat shock proteins regulated by heat shock factor-1.

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