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Articles in PresS, published online ahead of print February 6, 2002
Am J Physiol Cell Physiol, 10.1152/ajpcell.00511.2001
Submitted on October 24, 2001
Accepted on December 30, 2001
production via Egr-11
1 Nutrition, Case Western Reserve University, Cleveland, OH, USA
* To whom correspondence should be addressed. E-mail: len2{at}po.cwru.edu.
Lipopolysaccharide (LPS) is a potent activator of tumor necrosis factor (TNF) production by macrophages. LPS stimulates the phosphorylation of ERK1/2, as well as increases TNF mRNA and protein accumulation in RAW 264.7 murine macrophages. However, the role of ERK1/2 activation in mediating LPS-stimulated TNF production is not well understood. Inhibition of ERK1/2 activation with PD98059, an inhibitor of ERK1/2 activation, or over-expression of dominant negative ERK1/2 decreased LPS-induced TNF mRNA quantity, as well as secreted TNF peptide. LPS increased Egr-1 quantity in the nucleus; this response was blunted in cells treated with PD98059. LPS rapidly increased Egr-1 binding to the TNF promoter; the peak of binding activity (30 min) preceded maximal increases in TNF mRNA (45-60 min). Increased Egr-1 binding to the TNF promoter in response to LPS was decreased in cells pretreated with PD98059 or transfected with dominant negative ERK1/2. Using a CAT reporter gene linked to the Egr-1 promoter, we show that LPS increased Egr-1 promoter activity via an ERK1/2- dependent mechanism. These results delineate the role of ERK1/2 activation of Egr-1 activity in mediating LPS-induced increases in TNF mRNA expression in macrophages.
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