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J. Biol. Chem., Vol. 275, Issue 52, 41396-41404, December 29, 2000
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From the The sodium pump, Na,K-ATPase, is an important
protein for maintaining intracellular ion concentration, cellular
volume, and ion transport and is regulated both transcriptionally and
post-transcriptionally. We previously demonstrated that hyperoxia
increased Na,K-ATPase
Up-regulation of Na,K-ATPase
1 Transcription
by Hyperoxia Is Mediated by SP1/SP3 Binding*
§,,, and
University of Minnesota, Department of Medicine,
Minneapolis, Minnesota 55455 and the ¶ Department of
Biochemistry, State University of New York, Health Science Center,
Brooklyn, New York 11203-2012
1 gene expression in Madin-Darby
canine kidney (MDCK) cells. In this study, we identify a DNA element
necessary for up-regulation of the Na,K-ATPase 1
transcription by hyperoxia and evaluate the nuclear proteins
responsible for this up-regulation. Transient transfection experiments
in MDCK cells using sequential 5'-deletions of the rat Na,K-ATPase
1 promoter-luciferase fusion gene demonstrated promoter
activation by hyperoxia between 
102 and +151. The hyperoxia response
was localized to a 7-base pair region between 62 and 55, which
contained a GC-rich region consistent with a consensus sequence for the
SP1 family, that was sufficient for up-regulation by hyperoxia. This GC
element exhibited both basal and hyperoxia-induced promoter activity
and bound both transcription factors SP1 and SP3 in electrophoretic
mobility shift assays. In addition, electrophoretic mobility
shift assays demonstrated increased binding of SP1/SP3 in cells exposed
to hyperoxia while mutation of this element eliminated protein binding.
Other GC sites within the proximal promoter also demonstrated
up-regulation of transcription by hyperoxia, however, the site at 55
had higher affinity for SP proteins.
*
Some of the results contained in Table I were published in
abstract form as a supplement to Chest 116, 87S-88S (Aspen
Lung Conference, 1998).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.
Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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