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Smad3 is acetylated by p300/CBP to regulate its transactivation activity

Oncogene volume 26, pages 500–508 (2007)Cite this article

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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Author notes

  1. Y Inoue

    Present address: Research Institute, National Cancer Center, Tokyo, Japan

  2. K Abe

    Present address: Department of Biochemistry 1, Hamamatsu University School of Medicine, Hamamatsu, Japan

Authors and Affiliations

  1. Department of Molecular Health Sciences, Graduate School of Pharmaceutical Sciences, Nagoya City University, Mizuho, Nagoya, Japan

    Y Inoue, Y Itoh, K Abe, K Onozaki & H Hayashi

  2. Department of Molecular and Cellular Biology, Nagoya City University Graduate School of Medical Sciences, Aichi, Nagoya, Japan

    T Okamoto

  3. Center for Tsukuba Advanced Research Alliance, Institute of Applied Biochemistry, University of Tsukuba, Tsukuba, Ibaraki, Japan

    H Daitoku & A Fukamizu

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  1. Y Inoue
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  2. Y Itoh
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Correspondence to H Hayashi.

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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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