Abstract
The bHLH transcription factor Hand1 is essential for placentation and cardiac morphogenesis in the developing embryo. Here we implicate Hand1 as a molecular switch that determines whether a trophoblast stem cell continues to proliferate or commits to differentiation. We identify a novel interaction of Hand1 with a protein that contains an I-mfa (inhibitor of myogenic factor) domain that anchors Hand1 in the nucleolus where it negatively regulates Hand1 activity. In the trophoblast stem-cell line Rcho-1, nucleolar sequestration of Hand1 accompanies sustained cell proliferation and renewal, whereas release of Hand1 into the nucleus leads to its activation, thus committing cells to a differentiated giant-cell fate. Site-specific phosphorylation is required for nucleolar release of Hand1, for its dimerization and biological function, and this is mediated by the non-canonical polo-like kinase Plk4 (Sak). Sak is co-expressed in Rcho-1 cells, localizes to the nucleolus during G2 and phosphorylates Hand1 as a requirement for trophoblast stem-cell commitment to a giant-cell fate. This study defines a novel cellular mechanism for regulating Hand1 that is a crucial step in the stem-cell differentiation pathway.
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Acknowledgements
We thank Anthony Firulli, Vivek Mittal and Sabine Thebault, for generously providing plasmids, Jean-Michel Mesnard for the α-HIC antibody and Michael Soares and Satoshi Tanaka for generously providing the Rcho-1 and TS cell lines respectively. This research was supported by the British Heart Foundation.
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D.M.J.M carried out the functional cell-based characterization studies. C.A.R. and N.S. contributed to northern and western data and histological sections from Sak-null embryos. M.D.M.F.-V. peformed the yeast two-hybrid screen. COR collected and prepared Sak-null embryos. C.J.S. and J.W.D. provided Sak-null embryos and critical appraisal of Sak-null embryo data. P.R.R. carried out initial characterization studies and Sak-null embryo analyses, devised the functional studies and wrote the manuscript.
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Martindill, D., Risebro, C., Smart, N. et al. Nucleolar release of Hand1 acts as a molecular switch to determine cell fate. Nat Cell Biol 9, 1131–1141 (2007). https://doi.org/10.1038/ncb1633
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DOI: https://doi.org/10.1038/ncb1633
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