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(Received for publication, March 18, 1996, and in revised form, July 11, 1996)
From the Department of Pharmacological and Physiological Science,
St. Louis University School of Medicine,
St. Louis, Missouri 63104
Many parathyroid hormone (PTH)-mediated events in
osteoblasts are thought to require immediate early gene expression. PTH
induces the immediate early gene, c-fos, in this cell type
through a cAMP-dependent pathway. The present work
investigated the nuclear mechanisms involved in PTH regulation of
c-fos in the osteoblastic cell line, UMR 106-01. By
transiently transfecting c-fos promoter 5
Volume 271, Number 41,
Issue of October 11, 1996
pp. 25715-25721
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.
deletion
constructs into UMR cells, we demonstrated that PTH induction of the
c-fos promoter requires the major cAMP response element
(CRE). Point mutations created in the major CRE within the largest
construct inhibited both PTH-stimulated and basal expression. This
element, therefore, performs concerted basal and PTH-responsive
cis-acting functions. Gel retardation and Western blotting
techniques revealed that CRE-binding protein (CREB) constitutively
binds the major CRE but becomes phosphorylated at its
cAMP-dependent protein kinase consensus recognition site
following PTH treatment. CREB was functionally implicated in
c-fos regulation by coexpressing a dominant CREB repressor,
KCREB (killer CREB), with the c-fos promoter constructs.
KCREB suppressed both basal and PTH-mediated c-fos
induction. We conclude that PTH activates c-fos in
osteoblasts through cAMP-dependent protein
kinase-phosphorylated CREB interaction with the major CRE in the
promoter region of the c-fos gene.
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