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Endocrinology, Vol 136, 3901-3908, Copyright © 1995 by Endocrine Society
ARTICLES |
TL McCarthy, MJ Thomas, M Centrella and P Rotwein
Section of Plastic Surgery, Yale University School of Medicine, New Haven, Connecticut 06520-8041, USA.
Insulin-like growth factor I (IGF-I) is a locally synthesized anabolic growth factor for bone. IGF-I synthesis by primary fetal rat osteoblasts (Ob) is stimulated by agents that increase the intracellular cAMP concentration, including prostaglandin E2 (PGE2). Previous studies with Ob cultures demonstrated that PGE2 enhanced IGF-I transcription through selective use of IGF-I promoter 1, with little effect on IGF-I messenger RNA half-life. Transient transfection of Ob cultures with an array of promoter 1-luciferase reporter fusion constructs has now allowed localization of a potential cis-acting promoter element(s) responsible for cAMP-stimulated gene expression to the 5'-untranslated region (5'-UTR) of IGF-I exon 1, within a segment lacking a consensus cAMP response element. Our evidence derives from three principal observations: 1) a transfection construct containing only 122 nucleotides (nt) of promoter 1 and 328 nt of the 5'-UTR retained full PGE2-stimulated reporter expression; 2) maximal PGE2- driven reporter expression required the presence of nt 196 to 328 of exon 1 when tested within the context of IGF-I promoter 1; 3) cotransfection of IGF-I promoter-luciferase-reporter constructs with a plasmid encoding the alpha-isoform of the catalytic subunit of murine cAMP-dependent protein kinase (PKA) produced results comparable to those seen with PGE2 treatment, whereas cotransfection with a plasmid encoding a mutant regulatory subunit of PKA that cannot bind cAMP blocked PGE2-induced reporter expression. Deoxyribonuclease I footprinting of the 5'-UTR of exon 1 demonstrated protected sequences at HS3A, HS3B, and HS3D, three of six DNA-protein binding sites previously characterized with rat liver nuclear extracts. Of these three regions, only the HS3D binding site is located within the functionally identified hormonally responsive segment of IGF-I exon 1. These results directly implicate PKA in the control of IGF-I gene transcription by PGE2 and identify a segment of IGF-I exon 1 as being essential for this hormonal regulation.
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Y. Umayahara, J. Billiard, C. Ji, M. Centrella, T. L. McCarthy, and P. Rotwein CCAAT/Enhancer-binding Protein delta Is a Critical Regulator of Insulin-like Growth Factor-I Gene Transcription in Osteoblasts J. Biol. Chem., April 9, 1999; 274(15): 10609 - 10617. [Abstract] [Full Text] [PDF]
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Y. Umayahara, C. Ji, M. Centrella, P. Rotwein, and T. L. McCarthy CCAAT/Enhancer-binding Protein delta Activates Insulin-like Growth Factor-I Gene Transcription in Osteoblasts. IDENTIFICATION OF A NOVEL CYCLIC AMP SIGNALING PATHWAY IN BONE J. Biol. Chem., December 12, 1997; 272(50): 31793 - 31800. [Abstract] [Full Text] [PDF]
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P. Ye, Y. Umayahara, D. Ritter, T. Bunting, H. Auman, P. Rotwein, and A. J. D'Ercole Regulation of Insulin-Like Growth Factor I (IGF-I) Gene Expression in Brain of Transgenic Mice Expressing an IGF-I-Luciferase Fusion Gene Endocrinology, December 1, 1997; 138(12): 5466 - 5475. [Abstract] [Full Text] [PDF]
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J. H. Shin, C. Ji, S. Casinghino, T. L. McCarthy, and M. Centrella Parathyroid Hormone-related Protein Enhances Insulin-like Growth Factor-I Expression by Fetal Rat Dermal Fibroblasts J. Biol. Chem., September 19, 1997; 272(38): 23498 - 23502. [Abstract] [Full Text] [PDF]
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T. L. McCarthy, C. Ji, H. Shu, S. Casinghino, K. Crothers, P. Rotwein, and M. Centrella 17beta -Estradiol Potently Suppresses cAMP-induced Insulin-like Growth Factor-I Gene Activation in Primary Rat Osteoblast Cultures J. Biol. Chem., July 18, 1997; 272(29): 18132 - 18139. [Abstract] [Full Text] [PDF]
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X. Wang, Y. Yang, and M. L. Adamo Characterization of the Rat Insulin-Like Growth Factor I Gene Promoters and Identification of a Minimal Exon 2 Promoter Endocrinology, April 1, 1997; 138(4): 1528 - 1536. [Abstract] [Full Text] [PDF]
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M. J. Thomas, Y. Umayahara, H. Shu, M. Centrella, P. Rotwein, and T. L. McCarthy Identification of the cAMP Response Element That Controls Transcriptional Activation of the Insulin-like Growth Factor-I Gene by Prostaglandin E2 in Osteoblasts J. Biol. Chem., September 6, 1996; 271(36): 21835 - 21841. [Abstract] [Full Text] [PDF]
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T. L. McCarthy, S. Casinghino, D. W. Mittanck, C.-H. Ji, M. Centrella, and P. Rotwein Promoter-dependent and -independent Activation of Insulin-like Growth Factor Binding Protein-5 Gene Expression by Prostaglandin E(2) in Primary Rat Osteoblasts J. Biol. Chem., March 22, 1996; 271(12): 6666 - 6671. [Abstract] [Full Text] [PDF]
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T. L. McCarthy, C. Ji, Y. Chen, K. K. Kim, M. Imagawa, Y. Ito, and M. Centrella Runt Domain Factor (Runx)-dependent Effects on CCAAT/ Enhancer-binding Protein delta Expression and Activity in Osteoblasts J. Biol. Chem., July 7, 2000; 275(28): 21746 - 21753. [Abstract] [Full Text] [PDF]
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J. Billiard, Y. Umayahara, K. Wiren, M. Centrella, T. L. McCarthy, and P. Rotwein Regulated Nuclear-Cytoplasmic Localization of CCAAT/ Enhancer-binding Protein delta in Osteoblasts J. Biol. Chem., April 27, 2001; 276(18): 15354 - 15361. [Abstract] [Full Text] [PDF]
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J. Billiard, S. S. Grewal, L. Lukaesko, P. J. S. Stork, and P. Rotwein Hormonal Control of Insulin-like Growth Factor I Gene Transcription in Human Osteoblasts. DUAL ACTIONS OF cAMP-DEPENDENT PROTEIN KINASE ON CCAAT/ENHANCER-BINDING PROTEIN delta J. Biol. Chem., August 10, 2001; 276(33): 31238 - 31246. [Abstract] [Full Text] [PDF]
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