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J. Biol. Chem., Vol. 276, Issue 2, 1211-1219, January 12, 2001
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From the Our recent study indicates that
lysophosphatidylcholine (LPC) enhances Sp1 binding and
Sp1-dependent endothelial nitric oxide synthase (eNOS)
promoter activity via the mitogen-activated protein kinase/extracellular signal-regulated kinase kinase 1 (MEK-1) signaling pathway (Cieslik, K., Lee, C.-M., Tang, J.-L., and Wu, K. K. (1999) J. Biol. Chem. 274, 34669-34675).
To identify upstream signaling molecules, we transfected human
endothelial cells with dominant negative and active mutants of Ras and
evaluated their effects on eNOS promoter activity. Neither mutant
altered the basal or LPC-induced eNOS promoter function. By contrast, a
dominant negative mutant of phosphatidylinositol 3-kinase
Up-regulation of Endothelial Nitric-oxide Synthase Promoter by
the Phosphatidylinositol 3-Kinase
/Janus Kinase
2/MEK-1-dependent Pathway*
,¶
Vascular Biology Research Center and
Division of Hematology, University of Texas Medical School,
Houston, Texas 77030 and § Department of Medicine,
University of Pennsylvania, School of Medicine,
Philadelphia, Pennsylvania 19104
(PI-3K) blocked the promoter activity induced by LPC. Wortmannin and
LY 294002 had a similar effect. AG-490, a selective
inhibitor of Janus kinase 2 (Jak2), also reduced the LPC-induced
Sp1 binding and eNOS promoter activity to the basal level. LPC induced
Jak2 phosphorylation, which was abolished by LY 294002 and the dominant
negative mutant of PI-3K. LY 294002 and AG-490 abrogated MEK-1
phosphorylation induced by LPC but had no effect on Raf-1. These
results indicate that PI-3K and Jak2 are essential for LPC-induced
eNOS promoter activity. This signaling pathway was sensitive to
pertussis toxin, suggesting the involvement of a Gi protein
in PI-3K activation. These results indicate that LPC enhances
Sp1-dependent eNOS promoter activity by a pertussis
toxin-sensitive, Ras-independent novel pathway,
PI-3K/Jak2/MEK-1/ERK1/2.
*
This work was supported by National Institutes of Health
Grants P50 NS23327 and RO1 HL50675.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.
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