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Cyclic AMP enhances Smad-mediated BMP signaling through PKA-CREB pathway

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Abstract

We present experimental results indicating involvement of cyclic AMP (cAMP)-mediated signaling in bone morphogenetic protein (BMP)-induced osteoblastic gene expression at the transcriptional level by luciferase activity assay in C2C12 cells using the promoter sequence of the Id1 gene, an early-response gene to BMPs, which contains both a BMP-responsive element (BRE) and a cAMP-response element (CRE). In cells transfected with luciferase gene driven by wild-type Id1 promoter, treatment with BMP-4 increased luciferase expression, which was further enhanced by the addition of dibutyryl cAMP (dbcAMP). This dbcAMP-enhanced luciferase expression was significantly suppressed when the CRE site in the Id1 promoter was replaced by mutated CRE or endogenous CRE-binding protein (CREB) was knocked down by transfection of CREB RNAi. Pretreatment of cells with protein kinase A (PKA) inhibitor, H89, also dramatically reduced dbcAMP-enhanced luciferase expression. Immunoprecipitation assay showed phosphorylated-Smad1/5/8, phosphorylated-CREB, and CREB-binding protein (CBP) formed the transcriptional complex. These data indicate that cAMP-PKA/CREB/CRE signaling potentially enhances BMP-induced transcription through the BRE in the promoter of the BMP-responsive gene through a PKA-mediated pathway.

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Authors and Affiliations

  1. Department of Orthopaedic Surgery, Osaka City University Graduate School of Medicine, 1-4-3 Asahi-machi, Abeno-ku, Osaka, 545-8585, Japan

    Yoichi Ohta, Keisuke Nakagawa, Yuuki Imai, Tatsuya Koike & Kunio Takaoka

  2. Division of Pathophysiology, Research Center for Genomic Medicine, Saitama Medical School, Saitama, Japan

    Takenobu Katagiri

Authors
  1. Yoichi Ohta
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  2. Keisuke Nakagawa
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  3. Yuuki Imai
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  4. Takenobu Katagiri
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  5. Tatsuya Koike
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  6. Kunio Takaoka
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Correspondence to Yoichi Ohta.

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Ohta, Y., Nakagawa, K., Imai, Y. et al. Cyclic AMP enhances Smad-mediated BMP signaling through PKA-CREB pathway. J Bone Miner Metab 26, 478–484 (2008). https://doi.org/10.1007/s00774-008-0850-8

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  • DOI: https://doi.org/10.1007/s00774-008-0850-8

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