Expression of a Dominant-Negative Mutant of p21ras Inhibits Induction of Nitric Oxide Synthase and Activation of Nuclear Factor-kB in Primary Astrocytes

Authors

Kalipada Pahan, Department of Oral Biology, University of Nebraska Medical CenterFollow
Xiaojuan Lu, Department of Oral Biology, University of Nebraska Medical Center
Michael J. McKinney, Mayo Clinic
Charles Wood, University of Nebraska - LincolnFollow
Faruk G. Sheikh, Department of Biology, Walter Reed Army Institute of Research
John R. Raymond, Department of Medicine, Medical University of South Carolina

Document Type

Article

Date of this Version

2000

Citation

Journal of Neurochemistry, 2000

Comments

U.S. Government work

Abstract

The present study underlines the importance of p21ras in regulating the inducible nitric oxide synthase (iNOS) in primary astrocytes. Bacterial lipopolysaccharides induced the GTP loading of p21ras, and the expression of a dominant-negative mutant of p21ras (Dp21ras) inhibited lipopolysaccharide-induced GTP loading in rat primary astrocytes. To delineate the role of p21ras in the induction of iNOS, we examined the effect of Dp21ras on the expression of iNOS and the production of nitric oxide. It is interesting that expression of Dp21ras markedly inhibited the production of nitric oxide and the expression of iNOS in lipopolysaccharide- and pro-inflammatory cytokine (tumor necrosis factor-a, interleukin-1b; interferon- g)-stimulated rat and human primary astrocytes. Inhibition of iNOS promoter-derived chloramphenicol acetyltransferase activity by Dp21ras suggests that p21ras is involved in the transcription of iNOS. As activation of nuclear factor-kB (NF-kB) is necessary for the transcription of iNOS, we examined the effect of Dp21ras on the activation of NF-kB. Expression of Dp21ras inhibited the DNA binding as well as the transcriptional activity of NF-kB in activated astrocytes, suggesting that Dp21ras inhibits the expression of iNOS by inhibiting the activation of NF-kB. These studies also suggest that inhibitors of p21ras may be used as therapeutics in nitric oxide- and cytokine-mediated neuro-inflammatory diseases.