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Mechanism and function of heat shock-dependent IkappaBalpha expression.

K E Dunsmore¹, A G Denenberg, K Page

¹Division of Critical Care Medicine, Cincinnati Children's Hospital Medical Center and Cincinnati Children's Research Foundation, Cincinnati, OH 45229, USA.

Inflammation Research : Official Journal of the European Histamine Research Society ... [Et Al.]

|September 7, 2006

Summary

Heat shock enhances the stability of IkappaBalpha mRNA, leading to increased expression of this NF-kappaB inhibitor. This process, partly mediated by p38 MAP kinase, ultimately inhibits NF-kappaB activation.

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Area of Science:

Cellular stress response
Molecular biology
Immunology

Background:

Heat shock is known to inhibit the Nuclear Factor kappa B (NF-kappa B) pathway.
De novo expression of the intracellular NF-kappa B inhibitor, Ikappa Balpha, is a potential mechanism for this inhibition.

Purpose of the Study:

To elucidate the mechanisms by which heat shock induces Ikappa Balpha gene expression.
To determine the functional consequences of heat shock-mediated Ikappa Balpha gene expression in A549 cells.

Main Methods:

Nuclear run-on assays to assess Ikappa Balpha gene transcription.
Actinomycin D treatment to determine mRNA stability.
p38 MAP kinase activation and inhibition studies.
Tumor necrosis factor-alpha (TNF-alpha) stimulation assays.

Related Experiment Videos

Ikappa Balpha anti-sense oligonucleotide experiments.

Main Results:

Heat shock significantly increased the half-life of Ikappa Balpha mRNA, indicating enhanced mRNA stability.
Heat shock induced p38 MAP kinase activation, which was essential for heat shock-dependent Ikappa Balpha mRNA expression.
Following heat shock, NF-kappaB activation by TNF-alpha was inhibited, and this inhibition was reversed by Ikappa Balpha anti-sense oligonucleotides.

Conclusions:

Heat shock primarily increases Ikappa Balpha gene expression through enhanced mRNA stability, partially dependent on p38 MAP kinase.
The functional outcome of heat shock-induced Ikappa Balpha expression is the inhibition of NF-kappaB activation.