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Early B cell factor cooperates with Runx1 and mediates epigenetic changes associated with mb-1 transcription

Abstract

Cd79a (called mb-1 here) encodes the Ig-α signaling component of the B cell receptor. The early B cell–specific mb-1 promoter was hypermethylated at CpG dinucleotides in hematopoietic stem cells but became progressively unmethylated as B cell development proceeded. The transcription factor Pax5 activated endogenous mb-1 transcription in a plasmacytoma cell line, but could not when the promoter was methylated. In this context, early B cell factor (EBF), a transcription factor required for B lymphopoiesis, potentiated activation of mb-1 by Pax5. EBF and the basic helix-loop-helix transcription factor E47 each contributed to epigenetic modifications of the mb-1 promoter, including CpG demethylation and nucleosomal remodeling. EBF function was enhanced by interaction with the transcription factor Runx1. These data suggest a molecular basis for the hierarchical dependence of Pax5 function on EBF and E2A in B lymphocyte development.

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Figure 1: Sequencing results of sodium bisulfite–modified genomic DNA from sorted ex vivo bone marrow populations.
Figure 2: Runx proteins bind to mb-1 promoter SPR2 sites.
Figure 3: EBF and Runx1 cooperatively activate endogenous mb-1 transcription.
Figure 4: Reduced CpG methylation in μM.2 cells transduced to express EBF and sequencing results of sodium bisulfite-modified genomic DNA derived from μM.2 cells.
Figure 5: EBF activates nucleosomal remodeling of the mb-1 promoter.
Figure 6: EBF is required for hypomethylation of the distal mb-1 promoter.
Figure 7: EBF and E47 each enhance accessibility of mb-1 promoters in Tcfe2a−/− pre-pro–B cells.

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Acknowledgements

The authors thank P. Kincade (Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma) for providing the SB199 hybridoma; M. Reth (Max-Planck Institute, Tubingen, Germany) for providing the 558LμM cell line; M. Busslinger (Institute for Molecular Pathology, Vienna, Austria) for providing the Pax5−/− mice; and S. McNeff for assistance with illustrations. Supported by National Institutes of Health (T32 AI07405 to H.M.; R01 AI26782 and R01 AI40946 to R.R.H.; and P01 AI22295, R01 AI054661 and R01 AI056322 to J.H.), the Irvington Institute for Immunological Research (K.L.M.) and the Milheim Foundation.

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Correspondence to James Hagman.

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Supplementary information

Supplementary Fig. 1

EBF and Runx1/CBFβ synergistically activate the mb-1 promoter. (PDF 88 kb)

Supplementary Fig. 2

Real-time PCR analyses of Tcfe2a−/− pre-pro-B cells transduced with control, EBF or E47-expressing retroviruses. (PDF 128 kb)

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Maier, H., Ostraat, R., Gao, H. et al. Early B cell factor cooperates with Runx1 and mediates epigenetic changes associated with mb-1 transcription. Nat Immunol 5, 1069–1077 (2004). https://doi.org/10.1038/ni1119

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