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Epigenetic repression of the Igk locus by STAT5-mediated recruitment of the histone methyltransferase Ezh2

Nature Immunology volume 12, pages 1212–1220 (2011)Cite this article

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Abstract

During B lymphopoiesis, recombination of the locus encoding the immunoglobulin κ-chain complex (Igk) requires expression of the precursor to the B cell antigen receptor (pre-BCR) and escape from signaling via the interleukin 7 receptor (IL-7R). By activating the transcription factor STAT5, IL-7R signaling maintains proliferation and represses Igk germline transcription by unknown mechanisms. We demonstrate that a STAT5 tetramer bound the Igk intronic enhancer (Eκi), which led to recruitment of the histone methyltransferase Ezh2. Ezh2 marked trimethylation of histone H3 at Lys27 (H3K27me3) throughout the κ-chain joining region (Jκ) to the κ-chain constant region (Cκ). In the absence of Ezh2, IL-7 failed to repress Igk germline transcription. H3K27me3 modifications were lost after termination of IL-7R–STAT5 signaling, and the transcription factor E2A bound Eκi, which resulted in acquisition of H3K4me1 and acetylated histone H4 (H4Ac). Genome-wide analyses showed a STAT5 tetrameric binding motif associated with transcriptional repression. Our data demonstrate how IL-7R signaling represses Igk germline transcription and provide a general model for STAT5-mediated epigenetic transcriptional repression.

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Figure 1: Binding of STAT5 at κS2 in Eκi is functionally important.
Figure 2: STAT5 and E2A compete for binding at their respective Eκi sites.
Figure 3: Epigenetic regulation of Eκi during B lymphopoiesis.
Figure 4: STAT5 and E2A mediate repressive and activation marks, respectively, from Jκ through Cκ.
Figure 5: Binding of tetrameric STAT5 to κS2 recruits Ezh2 and represses Igk germline transcription.
Figure 6: Binding of tetrameric STAT5 mediates H3K27 trimethylation in vivo.
Figure 7: Trimethylation of H3K27 correlates with STAT5 target genes repressed throughout B cell development.

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Acknowledgements

We thank H. Singh and U. Storb for discussions; R. Duggan and D. Leclerc for cell-sorting services; and the ImmGen Consortium for data assembly. Supported by the US National Institutes of Health (GM088847 to M.R.C. and CA099978 to B.L.K.), the US Department of Energy (M.M.-C.), the Chicago National Institutes of Health Systems Biology Center (P50 GM081892 to A.R.D.) and the Leukemia and Lymphoma Society (B.L.K.).

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Authors and Affiliations

  1. Department of Medicine, Section of Rheumatology and Gwen Knapp Center for Lupus and Immunology Research, University of Chicago, Chicago, Illinois, USA

    Malay Mandal, Sarah E Powers, Keith M Hamel & Marcus R Clark

  2. Department of Chemistry and James Franck Institute, University of Chicago, Chicago, Illinois, USA

    Mark Maienschein-Cline & Aaron R Dinner

  3. University of Chicago Comprehensive Cancer Center, University of Chicago, Chicago, Illinois, USA

    Elizabeth T Bartom

  4. Department of Pathology, University of Chicago, Chicago, Illinois, USA

    Barbara L Kee

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Contributions

M.M. designed, did and analyzed most of the experiments and prepared the first draft of the paper; S.E.P. assisted in the design and analysis of many experiments; M.M.-C. compared mRNA expression and ChIP-Seq data, assisted by K.M.H.; E.T.B. assisted in the ChIP-Seq analysis. B.L.K. provided E2A-specific reagents and contributed to the design of some experiments; A.R.D. oversaw the analysis of microarray and ChIP-Seq data; and M.R.C. oversaw the entire project and prepared the final manuscript.

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Correspondence to Marcus R Clark.

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Mandal, M., Powers, S., Maienschein-Cline, M. et al. Epigenetic repression of the Igk locus by STAT5-mediated recruitment of the histone methyltransferase Ezh2. Nat Immunol 12, 1212–1220 (2011). https://doi.org/10.1038/ni.2136

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