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J Biol Chem, Vol. 274, Issue 48, 34301-34309, November 26, 1999
From the Department of Pathology, Division of Molecular and
Cellular Pathology, University of Alabama at Birmingham,
Birmingham, Alabama 35294-0019
Vascular smooth muscle cells (VSMC) exist in
either a contractile or a synthetic phenotype in vitro and
in vivo. The molecular mechanisms regulating phenotypic
modulation are unknown. Previous studies have suggested that the
serine/threonine protein kinase mediator of nitric oxide (NO) and
cyclic GMP (cGMP) signaling, the cGMP-dependent protein
kinase (PKG) promotes modulation to the contractile phenotype in
cultured rat aortic smooth muscle cells (RASMC). Because of the
potential importance of the mitogen-activated protein kinase (MAP
kinase) pathways in VSMC proliferation and phenotypic modulation, the
effects of PKG expression in PKG-deficient and PKG-expressing adult
RASMC on MAP kinases were examined. In PKG-expressing adult RASMC,
8-para-chlorophenylthio-cGMP activated extracellular
signal- regulated kinases (ERK1/2) and c-Jun N-terminal kinase (JNK).
The major effect of PKG activation was increased activation by MAP
kinase kinase (MEK). The cAMP analog, 8-Br-cAMP inhibited ERK1/2
activation in PKG-deficient and PKG-expressing RASMC but had no effect
on JNK activity. The effects of PKG on ERK and JNK activity were
additive with those of platelet-derived growth factor (PDGF),
suggesting that PKG activates MEK through a pathway not used by PDGF.
The stimulatory effects of cGMP on ERK and JNK activation were also
observed in low-passaged, contractile RASMC still expressing endogenous
PKG, suggesting that the effects of PKG expression were not artifacts
of cell transfections. These results suggest that in contractile adult
RASMC, NO-cGMP signaling increases MAP kinase activity. Increased
activation of these MAP kinase pathways may be one mechanism by which
cGMP and PKG activation mediate c-fos induction and
increased proliferation of contractile adult RASMC.
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