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J. Biol. Chem., Vol. 277, Issue 24, 21325-21331, June 14, 2002
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From the Previously we have demonstrated that activation
of p38 mitogen-activated protein kinase (MAPK) and induction of DNA
synthesis in response to receptor tyrosine kinase (RTK) and G
protein-coupled receptor (GPCR) agonists require NADH/NADPH-like
oxidase activity in vascular smooth muscle cells (VSMC). Here we tested
the role of p38 MAPK in RTK and GPCR agonist-induced DNA synthesis in
VSMC. Platelet-derived growth factor (PDGF)-BB and thrombin (RTK and GPCR agonists, respectively) activated p38 MAPK in a
time-dependent manner in VSMC. Inhibition of p38 MAPK led
to a 50% decrease in the DNA synthesis induced by thrombin but not
PDGF-BB. ATF-1 was found to be the predominant member of the cyclic AMP
response element (CRE)-DNA complex formed in VSMC in response to
PDGF-BB and thrombin, and both agonists induced its phosphorylation.
Regardless of this, inhibition of p38 MAPK reduced only thrombin- but
not PDGF-BB-induced ATF-1 phosphorylation. Similarly, inhibition of p38
MAPK caused a 50% decrease in thrombin- but not PDGF-BB-induced CRE
promoter-dependent transcription. Ectopic expression of an inhibitory anti-ATF-1 single-chain antibody fragment, ScFv,
significantly interfered with DNA synthesis induced by thrombin but not
PDGF-BB. Together, these results suggest the following conclusions. 1) Both RTK and GPCR agonists activate p38 MAPK and induce CRE
promoter-dependent transcription; 2) both RTK and GPCR
agonists induce ATF-1 phosphorylation, and ATF-1 is a predominant
member in the CRE-DNA complexes formed in response to these
agents; and 3) p38 MAPK-dependent ATF-1
phosphorylation and CRE promoter-mediated transcription are associated
with GPCR agonist-induced VSMC growth.
ATF-1 Mediates Protease-activated Receptor-1 but Not Receptor
Tyrosine Kinase-induced DNA Synthesis in Vascular Smooth Muscle
Cells*
§,§,,,
Department of Physiology, University of
Tennessee Health Science Center, Memphis, Tennessee 38163 and the
¶ Department of Cancer Biology, University of Texas M. D. Anderson Cancer Center, Houston, Texas 77030
*
This work was supported in part by grants from the National
Institutes of Health (to G. N. R.).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
Physiology, University of Tennessee Health Science Center, 894 Union Ave., Memphis, TN 38163. Tel.: 901-448-7321; Fax: 901-448-7126; E-mail:
grao@physio1.utmem.edu.
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