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Cascading suppression of transcriptional silencers by ThPOK seals helper T cell fate

Abstract

CD4 and the transcription factor ThPOK are essential for the differentiation of major histocompatibility complex class II–restricted thymocytes into the helper T cell lineage; their genes (Cd4 and Zbtb7b (called 'ThPOK' here)) are repressed by transcriptional silencer elements in cytotoxic T cells. The molecular mechanisms regulating expression of these genes during helper T cell lineage differentiation remain unknown. Here we showed that inefficient upregulation of ThPOK, induced by removal of the proximal enhancer from the ThPOK locus, resulted in the transdifferentiation of helper lineage–specified cells into the cytotoxic T cell lineage. Furthermore, direct antagonism by ThPOK of the Cd4 and ThPOK silencers generated two regulatory loops that initially inhibited Cd4 downregulation and later stabilized ThPOK expression. Our results show how an initial lineage–specification signal can be amplified and stabilized during the lineage–commitment process.

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Figure 1: CD8 lineage-differentiation potential of ThPOK-expressing CD4+CD8int thymocytes.
Figure 2: The ThPOK proximal enhancer is essential for efficient ThPOK upregulation.
Figure 3: The ThPOK proximal enhancer is essential for the normal development of CD4+ T cells.
Figure 4: Transdifferentiation of CD4+CD8 cells into CD4CD8 or CD4CD8+ cells.
Figure 5: Partial loss of helper identity and a partial acquisition of cytotoxic features by CD4+ T cells from ThPOKPEΔ/GFP mice.
Figure 6: ThPOK inhibits gene repression mediated by the Cd4 and ThPOK silencers in MHC class II–restricted cells.
Figure 7: Binding of ThPOK to silencers in the Cd4 and ThPOK loci antagonizes silencer activity.

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Acknowledgements

We thank D.J. Kappes (Fox Chase Cancer Center) for cDNA encoding helper-deficient mutant ThPOK; Y. Nakatani (Dana-Farber Cancer Institute) for the pOZ-FH-N vector; M. Matsuda for aggregation of embryonic stem cells; I. Hisanaga for microinjection of transgenes; H. Fujimoto and Y. Hachiman for cell sorting; and W. Ellmeier and S.L. Reiner for critical reading of the manuscript. Supported by Precursory Research for Embryonic Science and Technology, Japan Science and Technology Agency (I.T.).

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S.M. generated all mice with mutations at the ThPOK locus and did phenotypic analyses with assistance from C.M.; Y.N. did ChIP assays with tiling arrays with assistance from K.A.; T.I., K.M. and H.K. did in vitro culture in thymic lobes; and I.T. designed the experiments, interpreted the data and wrote the manuscript.

Corresponding author

Correspondence to Ichiro Taniuchi.

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Muroi, S., Naoe, Y., Miyamoto, C. et al. Cascading suppression of transcriptional silencers by ThPOK seals helper T cell fate. Nat Immunol 9, 1113–1121 (2008). https://doi.org/10.1038/ni.1650

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