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Mutations were made in (1) the cAMP response element (CRE) at \u221257\/\u221252\u2009bp, (2) the activating protein-1 (AP-1)\u2013binding site at \u221269\/\u221263\u2009bp, (3) the nuclear factor of activated T-cells (NFAT)\u2013binding site at \u221277\/\u221273\u2009bp, and (4) both the AP-1 and NFAT sites, which comprise a composite consensus sequence for NFAT\/AP-1. Single mutation of CRE, AP-1, or NFAT sites decreased parathyroid hormone (PTH)\u2013stimulated COX-2 promoter activity 40% to 60%, whereas joint mutation of NFAT and AP-1 abrogated the induction. On electrophoretic mobility shift analysis, PTH stimulated binding of phosphorylated CREB to an oligonucleotide spanning the CRE and binding of NFATc1, c-Fos, and c-Jun to an oligonucleotide spanning the NFAT\/AP-1 composite site. Mutation of the NFAT site was less effective than mutation of the AP-1 site in competing binding to the composite element, suggesting that cooperative interactions of NFATc1 and AP-1 are more dependent on NFAT than on AP-1. Both PTH and forskolin, an activator of adenylyl cyclase, stimulated NFATc1 nuclear translocation. PTH- and forskolin-stimulated COX-2 promoter activity was inhibited 56% to 80% by calcium chelation or calcineurin inhibitors and 60% to 98% by protein kinase A (PKA) inhibitors. These results indicate an important role for the calcium-calcineurin-NFAT signaling pathway in the PTH induction of COX-2 and suggest that cross-talk between the cAMP\/PKA pathway and the calcium-calcineurin-NFAT pathway may play a role in other functions of PTH in osteoblasts. \u00a9 2010 American Society for Bone and Mineral Research<\/jats:p>","DOI":"10.1359\/jbmr.091019","type":"journal-article","created":{"date-parts":[[2009,10,12]],"date-time":"2009-10-12T22:34:50Z","timestamp":1255386890000},"page":"819-829","source":"Crossref","is-referenced-by-count":22,"title":["Parathyroid hormone induction of cyclooxygenase-2 in murine osteoblasts: Role of the calcium-calcineurin-NFAT pathway"],"prefix":"10.1093","volume":"25","author":[{"given":"Hechang","family":"Huang","sequence":"first","affiliation":[{"name":"Department of Medicine, University of Connecticut Health Center, Farmington, CT, 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