Abstract
Id1 is a member of the inhibitor of differentiation (Id) protein family that regulates a wide range of cell functions. Previous studies have shown that expression of the Id1 gene is down-regulated by TGF-β in epithelial cells, whereas it is up-regulated by BMP in a variety of cell types. During our study of the biological function of TGF-β1, we found that Id1 can be strongly up-regulated by TGF-β1 in the human mammary gland epithelial cell line MCF10A. Quantitative real-time RT-PCR has revealed as high as 7.5-fold induction of Id1 mRNA by TGF-β1 in MCF10A cells after 1 h of TGF-β1 stimulation, and this induction does not require de novo protein synthesis. Using Smad knockdown and knockout approaches, we have identified Smad3 as the responsible R-Smad for mediating transcriptional activation of the Id1 gene. Chromatin immunoprecipitation assay confirms that Smad3 and Smad4 bind to the upstream region of the Id1 gene. Our results demonstrate that Smad3, but not Smad2, mediates TGF-β1-dependent early transcriptional induction of Id1.
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Acknowledgements
We thank Robert Benezra (Sloan-Kettering Institute, NY, USA) for Id1-luc reporter constructs and Ed Leof (Mayo Clinic, MN, USA) for anti-phospho-Smad3 antibody. We are grateful to Xin-Hua Feng (Baylor College of Medicine, TX, USA)for critically reading the manuscript and stimulating discussions. This research was supported by NIH grant R01DK073932, USA and partly by the Baylor Breast Center SPORE career development award, USA.
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Liang, YY., Brunicardi, F. & Lin, X. Smad3 mediates immediate early induction of Id1 by TGF-β. Cell Res 19, 140–148 (2009). https://doi.org/10.1038/cr.2008.321
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DOI: https://doi.org/10.1038/cr.2008.321
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