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J. Biol. Chem., Vol. 277, Issue 46, 44485-44496, November 15, 2002
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From the Divisions of The expression of the matrix cytokine osteopontin
(OPN) is up-regulated in aortic vascular smooth muscle cells (VSMCs) by diabetes. OPN expression in cultured VSMCs is reciprocally
regulated by glucose and 2-deoxyglucose (2-DG; inhibitor of cellular
glucose metabolism). Systematic analyses of OPN
promoter-luciferase reporter constructs identify a CCTCATGAC
motif at nucleotides
Osteopontin Transcription in Aortic Vascular Smooth Muscle Cells
Is Controlled by Glucose-regulated Upstream Stimulatory Factor and
Activator Protein-1 Activities*
§,,,,§¶
Bone and Mineral Diseases and
§ Endocrinology, Diabetes, and Metabolism, Department of
Internal Medicine, Washington University School of Medicine, St. Louis,
Missouri 63110
80 to 72 relative to the initiation site that
supports OPN transcription in VSMCs. The region 83 to
45 encompassing this motif confers basal and glucose- and
2-DG-dependent transcription on an unresponsive promoter.
Competition and gel mobility supershift assays identify upstream
stimulatory factor (USF; USF1:USF2) and activator protein-1 (AP1;
c-Fos:c-Jun) in complexes binding the composite CCTCATGAC element.
Glucose up-regulates both AP1 and USF binding activities 2-fold in A7r5
cells and selectively up-regulates USF1 protein levels. By contrast,
USF (but not AP1) binding activity is suppressed by 2-DG and restored
by glucose treatment. Expression of either USF or AP1 activates the
proximal OPN promoter in A7r5 VSMCs in part via the
CCTCATGAC element. Moreover, glucose stimulates the transactivation
functions of c-Fos and USF1, but not c-Jun, in one-hybrid assays.
Mannitol does not regulate binding, transactivation functions, USF1
protein accumulation, or OPN transcription. Thus, OPN gene transcription is regulated by USF and AP1 in aortic VSMCs, entrained to changes in cellular glucose metabolism.
*
This work was supported by Grants HL69229 and DK52446 (to
D. A. T.) from the National Institutes of Health and by the
Barnes-Jewish Hospital Research Foundation.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 © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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