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J Biol Chem, Vol. 275, Issue 4, 2811-2816, January 28, 2000
From the Department of Pharmacology, MC868, University of Illinois
at Chicago, Chicago, Illinois 60612
An increase in cellular levels of cyclic
nucleotides activates serine/threonine-dependent kinases
that lead to diverse physiological effects. Recently we reported the
activation of the p38 mitogen-activated protein kinase (MAPK) pathway
in neutrophils by a cGMP-dependent mechanism. In this study
we demonstrated that exogenously supplied nitric oxide leads to
activation of p38 MAPK in 293T fibroblasts. Phosphorylation of p38
corresponded with an increase in ATF-2-dependent gene
expression. The effect of nitric oxide was mimicked by addition of
8-bromo-cGMP, indicating that activation of soluble guanylyl cyclase
was involved. The importance of cGMP-dependent protein kinase in the activation of p38 MAPK by nitric oxide in 293T cells was
assessed in a transfection based assay. Overexpression of cGMP-dependent protein kinase-1
Nitric Oxide Activation of p38 Mitogen-activated Protein
Kinase in 293T Fibroblasts Requires cGMP-dependent
Protein Kinase*
,
caused phosphorylation
of p38 in these cells and potentiated the effectiveness of cGMP.
Overexpression of a catalytically inactive mutant form of this enzyme
(T516A) blocked the ability of both nitric oxide and 8-bromo-cGMP to
activate p38 as measured by both p38 phosphorylation and ATF-2 driven
gene expression. Together, these data demonstrate that nitric oxide stimulates a novel pathway leading to activation of p38 MAPK that requires activation of cGMP-dependent protein kinase.
*
This work was supported in part by National Institutes of
Health Grants AI33503 and AI40176.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.
Recipient of a fellowship from the Arthritis Foundation.
§
Established Investigator of the American Heart Association. To whom
correspondence should be addressed: Dept. of Pharmacology (MC868),
University of Illinois, 835 S. Wolcott Ave., Chicago, IL 60612. Tel.:
312-996-5087; Fax: 312-996-7857.
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