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J. Biol. Chem., Vol. 277, Issue 35, 31863-31870, August 30, 2002
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From the Department of Medicine, Division of Pulmonary and Critical
Care Medicine, Long Island Jewish Medical Center, Long Island Campus
for the Albert Einstein College of Medicine,
New Hyde Park, New York 11040
We examined the in vivo effect of
lipopolysaccharide (LPS) on Sp1 (promoter-selective transcription
factor 1) DNA binding activity and studied the mechanisms involved in
mouse lungs. The Sp1 DNA complex displayed a major band composed of
Sp1, Sp2, and Sp3 trimer and a minor band composed of Sp3 homodimer.
Compared with control, nuclear proteins from lungs challenged with LPS for 60, 90, 120, 150, 180, and 240 min, respectively, showed a markedly
reduced Sp1 binding activity. Down-regulation of Sp1 binding activity
was accompanied by a reduced expression of two Sp1-dependent genes (endothelial nitric oxide synthase and
cyclooxygenase-1). Immunoprecipitation-Western blot experiments
demonstrated that LPS dephosphorylated Sp1 protein at serine and
threonine residues but not at the tyrosine residue.
Dephosphorylation of Sp1 protein in vitro significantly
reduced Sp1 DNA binding activity. Deglycosylation of Sp1 protein also
reduced Sp1 binding activity. However, LPS did not cause Sp1
deglycosylation. LPS markedly reduced nuclear Sp1 protein level but had
no significant effect on Sp1 mRNA abundance and on Sp1 protein
nuclear translocation. Both Sp1 protein dephosphorylation and Sp1
protein degradation are temporally correlated to the reduced Sp1
binding activity. Our results demonstrate that challenge of mice with
LPS in vivo down-regulates Sp1 DNA binding activity through
promoting Sp1 protein dephosphorylation and degradation.
Lipopolysaccharide Down-regulates Sp1 Binding Activity by
Promoting Sp1 Protein Dephosphorylation and Degradation*
*
This work was supported in part by the North Shore LIJ
Research Institute Faculty Award Program.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: Division of Pulmonary
and Critical Care Medicine (RM C-20), Long Island Jewish Medical
Center, New Hyde Park, NY 11040. Tel.: 718-470-7253; Fax: 718-470-1507;
E-mail: Sliu@lij.edu.
Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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