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Originally published In Press as doi:10.1074/jbc.M204994200 on June 4, 2002

J. Biol. Chem., Vol. 277, Issue 33, 29584-29592, August 16, 2002
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p38 MAPK-mediated Transcriptional Activation of Inducible Nitric-oxide Synthase in Glial Cells
ROLES OF NUCLEAR FACTORS, NUCLEAR FACTOR kappa B, cAMP RESPONSE ELEMENT-BINDING PROTEIN, CCAAT/ENHANCER-BINDING PROTEIN-beta , AND ACTIVATING TRANSCRIPTION FACTOR-2*

Narayan R. BhatDagger §, Douglas L. Feinstein, Qin ShenDagger , and Aruna N. BhatDagger

From the Dagger  Department of Neurology, Medical University of South Carolina, Charleston, South Carolina 29425 and the  Department of Anesthesiology, University of Illinois, Chicago, Illinois 60612

Previous studies have shown that mitogen-activated protein kinase (MAPK) cascades signal the induction of inducible nitric-oxide synthase (iNOS) in glial cells (Bhat, N. R., Zhang, P., Lee, J. C., and Hogan E. L. (1998) J. Neurosci. 18, 1633-1641; Bhat, N. R., Zhang, P., and Bhat, A. N. (1999) J. Neurochem. 72, 472-478). This study further investigates the role of p38 MAPK in the transcriptional activation of the iNOS gene by transient transfection with constitutively active upstream kinases in the pathway (i.e. MAPK kinase 3 (MKK3b(E)) and MAPK kinase 6 (MKK6b(E)). Expression in C-6 glial cells of either MKK3b(E) or MKK6b(E) resulted in an induction of the activity of a cotransfected rat iNOS promoter-reporter (iNOS-luciferase (Luc)) gene and an enhancement of cytokine-induced expression of iNOS mRNA, both of which were inhibitable by the p38 MAPK inhibitor SB203580. The MKK constructs also induced cAMP response element-mediated (CRE-Luc) and nuclear factor kappa B-dependent (nuclear factor kappa B-Luc) transcriptional activities. Transfection with dominant negative (dn) forms of CRE-binding protein (CREB) and CCAAT/enhancer-binding protein (C/EBP), the two CRE-binding transcription factors targeted by the p38 MAPK pathway, resulted in opposite effects; dnCREB enhanced and dnC/EBP inhibited iNOS-Luc parallel to their effects on CRE-Luc. In addition, the induction, by MKK3b(E) and MKK6b(E), of iNOS promoter activity was enhanced by a wild-type activating transcription factor (ATF-2), whereas a phosphorylation-defective form of ATF-2 had a suppressive effect. The results of these molecular studies provide evidence for an important role for the p38 MAPK pathway in the transcriptional activation of the iNOS gene in rat glial cells involving the transcription factors nuclear factor kappa B, C/EBP, and ATF-2.

* This work was supported by National Institutes of Health Grants NS41035 (to N. R. B.) and NS31556 (to D. L. F.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

§ To whom correspondence should be addressed: Dept. of Neurology, Medical University of South Carolina, 171 Ashley Ave., Charleston, SC 29425. Tel.: 843-792-7593; Fax: 843-792-8626; E-mail: bhatnr@musc.edu.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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