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Transcription factors T-bet and Runx3 cooperate to activate Ifng and silence Il4 in T helper type 1 cells

Nature Immunology volume 8, pages 145–153 (2007)Cite this article

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Abstract

Cell differentiation involves activation and silencing of lineage-specific genes. Here we show that the transcription factor Runx3 is induced in T helper type 1 (TH1) cells in a T-bet-dependent manner, and that both transcription factors T-bet and Runx3 are required for maximal production of interferon-γ (IFN-γ) and silencing of the gene encoding interleukin 4 (Il4) in TH1 cells. T-bet does not repress Il4 in Runx3-deficient TH2 cells, but coexpression of Runx3 and T-bet induces potent repression in those cells. Both T-bet and Runx3 bind to the Ifng promoter and the Il4 silencer, and deletion of the silencer decreases the sensitivity of Il4 to repression by either factor. Our data indicate that cytokine gene expression in TH1 cells may be controlled by a feed-forward regulatory circuit in which T-bet induces Runx3 and then 'partners' with Runx3 to direct lineage-specific gene activation and silencing.

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Figure 1: Expression of Runx in peripheral T cells.
Figure 2: Runx3 contributes to activation of Ifng.
Figure 3: Runx3 binds to the Ifng promoter.
Figure 4: Runx3-deficient TH1 cells aberrantly express IL-4.
Figure 5: T-bet and Runx3 cooperate in repression of IL-4.
Figure 6: The Il4 silencer contributes to Runx3- and T-bet-mediated repression of IL-4.
Figure 7: T-bet and Runx3 interact with each other and the Il4 silencer.

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Change history

  • 17 January 2007

    In the version of this article initially published online, two labels in Figure 1e are reversed and two sequences in Figure 7a are reversed. The labels ‘IB: Runx3’ and ‘IB: T-bet’ should be switched, and the sequences for ‘HS IV probe’ and ‘T-box mutant’ should be switched. The errors have been corrected for all versions of the article.

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Acknowledgements

We thank L. Glimcher for Tbx21−/− mice, polyclonal antibody to T-bet, and Runx1 and Runx3 pCDNA3 expression plasmids; R. Locksley for DO11.10 Tcra,Tcrb transgenic Tcra−/− mice; K. Murphy for the parent retroviral vector GFP-RV; V. Heissmeyer for the pMSCV-IRES-Thy1.1 vector; G.P. Nolan for Phoenix cells; D. Bolton and L. Smith for technical support; J. Chermesh and R. Saka for help with animal husbandry; and F. Cruz-Guilloty and M. Keir for advice. Supported by the National Institutes of Health (AI44432 and HL67664 to A.R.), Sandler Program for Asthma Research (A.R.), The Leukemia and Lymphoma Society (K.M.A.) and the Israel Science Foundation (Y.G. and D.L.).

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Authors and Affiliations

  1. Harvard Medical School and the CBR Institute for Biomedical Research, Boston, 02115, Massachusetts, USA

    Ivana M Djuretic, Anjana Rao & K Mark Ansel

  2. Department of Molecular Genetics, The Weizmann Institute of Science, Rehovot, 76100, Israel

    Ditsa Levanon, Varda Negreanu & Yoram Groner

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  1. Ivana M Djuretic
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  2. Ditsa Levanon
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Contributions

I.M.D. did all the experiments with the direction and supervision of K.M.A. and A.R.; and D.L. and V.N. assisted in the analysis of Runx3-deficient T cells under the supervision of Y.G.

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Correspondence to Anjana Rao or K Mark Ansel.

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Supplementary information

Supplementary Fig. 1

Runx1 and Tbx21 mRNA expression. (PDF 971 kb)

Supplementary Fig. 2

Runx3 regulates IL-2 production. (PDF 371 kb)

Supplementary Fig. 3

Runx1 can bind the Ifng promoter and Il4 silencer. (PDF 302 kb)

Supplementary Fig. 4

Quantification of HS IV-dependent Runx3- and T-bet-mediated IL-4 repression. (PDF 372 kb)

Supplementary Fig. 5

Conserved T-box and Runx consensus binding sequences in HS IV. (PDF 469 kb)

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Djuretic, I., Levanon, D., Negreanu, V. et al. Transcription factors T-bet and Runx3 cooperate to activate Ifng and silence Il4 in T helper type 1 cells. Nat Immunol 8, 145–153 (2007). https://doi.org/10.1038/ni1424

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