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Originally published In Press as doi:10.1074/jbc.M006564200 on January 2, 2001

J. Biol. Chem., Vol. 276, Issue 15, 11639-11647, April 13, 2001
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PTHrP Modulates Chondrocyte Differentiation through AP-1 and CREB Signaling*

Andreia M. IonescuDagger , Edward M. Schwarz§, Charles Vinson, J. Edward Puzas§, Randy Rosier§, Paul R. Reynolds§, and Regis J. O'Keefe§||

From the Departments of Dagger  Biochemistry and Biophysics and the § Department of Orthopaedics, University of Rochester, School of Medicine and Dentistry, Rochester, New York 14642 and the  NCI, National Institutes of Health, Bethesda, Maryland 20892

During the process of differentiation, chondrocytes integrate a complex array of signals from local or systemic factors like parathyroid hormone-related peptide (PTHrP), Indian hedgehog, bone morphogenetic proteins and transforming growth factor beta . While PTHrP is known to be a critical regulator of chondrocyte proliferation and differentiation, the signaling pathways through which this factor acts remain to be elucidated. Here we show that both cAMP response element-binding protein (CREB) and AP-1 activation are critical to PTHrP signaling in chondrocytes. PTHrP treatment leads to rapid CREB phosphorylation and activation, while CREB DNA binding activity is constitutive. In contrast, PTHrP induces AP-1 DNA binding activity through induction of c-Fos protein expression. PTHrP activates CRE and TRE reporter constructs primarily through PKA-mediated signaling events. Both signaling pathways were found to be important mediators of PTHrP effects on chondrocyte phenotype. Alone, PTHrP suppresses maturation and stimulates proliferation of the chondrocyte cultures. However, in the presence of dominant negative inhibitors of CREB and c-Fos, these PTHrP effects were suppressed, and chondrocyte maturation was accelerated. Moreover, in combination, the effects of dominant negative c-Fos and CREB are synergistic, suggesting interaction between these signaling pathways during chondrocyte differentiation.

* This work was supported by National Institute of Arthritis, Musculoskeletal and Skin Disease Grant RO1 AR 38945 (to R. O'K.).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: Box 665, Dept. of Orthopaedics, University of Rochester Medical Center, 601 Elmwood Ave., Rochester, NY 14642. Tel.: 716-275-3100; Fax: 716-756-4727; E-mail: regis_okeefe@urmc.rochester.edu.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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