Abstract
In embryonic stem cells (ESCs), Wnt-responsive development-related genes are silenced to maintain pluripotency and their expression is activated during differentiation. Acetylation of histones by histone acetyltransferases stimulates transcription, whereas deacetylation of histones by HDACs is correlated with transcriptional repression. Although Wnt-mediated gene transcription has been intimately linked to the acetylation or deacetylation of histones, how Wnt signaling regulates this type of histone modification is poorly understood. Here, we report that Smek, a regulatory subunit of protein phosphatase 4 (PP4) complex, plays an important role in histone deacetylation and silencing of the Wnt-responsive gene, brachyury, in ESCs. Smek mediates recruitment of PP4c and HDAC1 to the Tcf/Lef binding site of the brachyury promoter and inhibits brachyury expression in ESCs. Activation of Wnt signaling during differentiation causes disruption of the Smek/PP4c/HDAC1 complex, resulting in an increase in histones H3 and H4 acetylation at the brachyury gene locus. These results suggest that the Smek-containing PP4 complex represses transcription of Wnt-responsive development-related genes through histone deacetylation, and that this complex is essential for ESC pluripotency maintenance.
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Acknowledgements
The authors would like to thank people from the Lu lab for constructive discussions. Jungmook Lyu was supported by a postdoctoral fellowship from CIRM. This work was partially supported by the CIRM Basic Biology Award (RB1-01353-1) and a grant from NIH (1R01NS067213-01A1).
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( Supplementary information is linked to the online version of the paper on the Cell Research website.)
Supplementary information
Supplementary information, Figure S1
Overexpression of Smek reduces differentiation of ESCs into mesoderm. (PDF 53 kb)
Supplementary information, Figure S2
Depletion of Smek inhibits neuroectoderm differentiation from ESCs. (PDF 117 kb)
Supplementary information, Figure S3
Localization of β-catenin in ESCs knocked down for Smek1 and Smek1. (PDF 172 kb)
Supplementary information, Figure S4
Occupancy of Smek at Tcf/Lef binding site region of islet1 and pitx2. (PDF 57 kb)
Supplementary information, Figure S5
Smek increases binding of HDAC1 to Tcf protein. (PDF 81 kb)
Supplementary information, Figure S6
The knockdown for PP4c induces differentiation of ESCs. (PDF 112 kb)
Supplementary information, Figure S7
Smek-mediated blockade of transcriptional activation by Wnt/β-catenin signaling is dependent on the dose of Wnt3a protein. (PDF 72 kb)
Supplementary information, Figure S8
β-catenin-independent Wnt signaling increases brachyury expression. (PDF 59 kb)
Supplementary information, Data S1
Materials and Methods (PDF 70 kb)
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Lyu, J., Jho, Eh. & Lu, W. Smek promotes histone deacetylation to suppress transcription of Wnt target gene brachyury in pluripotent embryonic stem cells. Cell Res 21, 911–921 (2011). https://doi.org/10.1038/cr.2011.47
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DOI: https://doi.org/10.1038/cr.2011.47
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