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Insulin-like growth factor binding protein-6 interacts with the thyroid hormone receptor α1 and modulates the thyroid hormone-response in osteoblastic differentiation

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Abstract

Insulin-like growth factor binding protein-6 (IGFBP-6) is a member of the insulin-like growth factor binding protein family, which has both Insulin-like growth factor-dependent and independent effects on cell growth. In previous studies, we have shown that recombinant IGFBP-6 could be translocated into the cell nucleus. But the effect in the nucleus of IGFBP-6 is not clear. In the present study, we use multiple methodologies including Glutathione S-transferase pull-down assay, co-immunoprecipitation, fluorescence resonance energy transfer to demonstrate that IGFBP-6 can directly interact with thyroid hormone receptor alpha 1 (TRα1) in vitro and in vivo. We also demonstrate that the DNA-binding domains and Ligand-binding domains of TRα1 and N-terminal domains and C-terminal domains of IGFBP-6 are involved in the interaction. This interaction also can block the formation of TR: retinoid X receptor heterodimers. Furthermore, immunofluorescence co-localization studies show IGFBP-6 and TRα1 could co-localize in the nucleus of the cells. Reporter gene experiment shows that IGFBP-6 negatively regulates the growth hormone promoter activity induced by ligand activated TRα1. Moreover, real-time RT-PCR demonstrates that IGFBP-6 could inhibit the osteocalcin mRNA transcription induced by Triiodothyronine (3,3′,5-Triiodo-L-thyronine, T3) in osteoblastic cells. Finally, alkaline phosphatase activity was significantly decreased in osteoblastic cells when the cells were transfected with IGFBP-6 in the presence of T3. In conclusion, these studies provide evidence that overexpression of IGFBP-6 suppresses osteoblastic differentiation regulated by TR in the present of T3.

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Acknowledgments

The authors thank Prof. David J. Mangelsdorf (Howard Hughes Medical Institute, UT Southwestern Medical Center, Department of Pharmacology) for giving us pGEX-2T-hRXRa. The authors also thank Prof. Norman Eberhardt (Mayo Clinic, Department of Medicine/Division of Endocrinology) and Dr. Peter A. Cattini (Department of Physiology, University of Manitoba) for giving them pGL3-GH promoter plasmid and other GH relative luciferase reporter plasmids.

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Authors and Affiliations

  1. National Laboratory of Medical Molecular Biology, Department of Biochemistry and Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences & Peking Union Medical College, 5 Dong Dan San Tiao St., 100005, Beijing, China

    Jia Qiu, Xiao-Li Ma, Xin Wang, Hong Chen & Bing-Ren Huang

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  1. Jia Qiu
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  2. Xiao-Li Ma
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  3. Xin Wang
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  4. Hong Chen
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Correspondence to Hong Chen or Bing-Ren Huang.

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Qiu, J., Ma, XL., Wang, X. et al. Insulin-like growth factor binding protein-6 interacts with the thyroid hormone receptor α1 and modulates the thyroid hormone-response in osteoblastic differentiation. Mol Cell Biochem 361, 197–208 (2012). https://doi.org/10.1007/s11010-011-1104-y

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  • DOI: https://doi.org/10.1007/s11010-011-1104-y

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