Abstract
Smad proteins are crucial for the intracellular signaling of transforming growth factor-β (TGF-β). Upon their receptor-induced activation, Smad proteins are phosphorylated and translocated to the nucleus to activate the transcription of a select set of target genes. Here, we show that the co-activator p300/CBP bound and acetylated Smad3 as well as Smad2 in vivo, and that the acetylation was stimulated by TGF-β. A major acetylation site of Smad3 by p300/CBP is Lys-378 in the MH2 domain (Smad3C) known to be critical for the regulation of transcriptional activity. Replacement of Lys-378 with Arg decreased the transcriptional activity of GAL4-Smad3C in a luciferase assay. Moreover, p300/CBP potentiated the transcriptional activity of GAL4-Smad3C, but not the acetylation-resistant GAL4-Smad3C(K378R) mutant. These results suggest that acetylation of Smad3 by p300/CBP regulates positively its transcriptional activity.
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Abbreviations
- BMP:
-
bone morphogenetic protein
- DMEM:
-
Dulbecco's modified Eagle's medium
- EDTA:
-
ethylenediamine tetraacetic acid
- FBS:
-
fetal bovine serum
- MH:
-
Mad homology
- PAGE:
-
polyacrylamide gel electrophoresis
- PCR:
-
polymerase chain reaction
- SDS:
-
sodium dodecyl sulfate
- TGF-β:
-
transforming growth factor-β
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Acknowledgements
We thank Dr JL Wrana (Samuel Lunenfeld Research Institute) and Dr K Miyazono (University of Tokyo) for providing plasmids. We also thank Dr C Uchida (Hamamatsu University) for kind suggestions. This work was supported in part by Grants-in-Aid for Scientific Research (C) from the Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research on Priority Areas (C) from the Ministry of Education, Science Sports and Culture, Welfide Medical Research Foundation, Grants-in-Aid for Scientific Research from Nagoya City University and the Kudo Research Foundation.
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Inoue, Y., Itoh, Y., Abe, K. et al. Smad3 is acetylated by p300/CBP to regulate its transactivation activity. Oncogene 26, 500–508 (2007). https://doi.org/10.1038/sj.onc.1209826
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DOI: https://doi.org/10.1038/sj.onc.1209826
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