Abstract
Tight regulation of transcription factors, such as PU.1, is crucial for generation of all hematopoietic lineages. We previously reported that mice with a deletion of an upstream regulatory element (URE) of the gene encoding PU.1 (Sfpi1) developed acute myeloid leukemia. Here we show that the URE has an essential role in orchestrating the dynamic PU.1 expression pattern required for lymphoid development and tumor suppression. URE deletion ablated B2 cells but stimulated growth of B1 cells in mice. The URE was a PU.1 enhancer in B cells but a repressor in T cell precursors. TCF transcription factors coordinated this repressor function and linked PU.1 to Wnt signaling. Failure of appropriate PU.1 repression in T cell progenitors with URE deletion disrupted differentiation and induced thymic transformation. Genome-wide DNA methylation assessment showed that epigenetic silencing of selective tumor suppressor genes completed PU.1-initiated transformation of lymphoid progenitors with URE deletion. These results elucidate how a single transcription factor, PU.1, through the cell context–specific activity of a key cis-regulatory element, affects the development of multiple cell lineages and can induce cancer.
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Acknowledgements
We thank K. Martens and K. Geary for their assistance in animal husbandry; T. Dayaram and C. Hetherington for help with the real time PCRs; Y.Z. Wu for help with RLGS analysis; J. Tigges and V. Toxavidis for assistance with multicolor flow cytometry; H. Clevers, R. Grosschedl, T. Reya, H. Singh, G. Nolan and J.C. Zuniga-Pflucker for reagents; and H. Clevers, M. Ye, T. Graf, A. Ebralidze and other members of the laboratory of D.G.T. for discussions and sharing of unpublished information. This work was supported by National Institutes of Health grants to T.L.R., C.P. and D.G.T. and by fellowships from the Lymphoma Research Foundation to F.R., from the American Cancer Society to B.M.O. and from the Dr. Mildred Scheel Foundation for Cancer Research to U.S. and B.H. Immunohistochemical methodology was supported by the Specialized Histopathology Core Lab of the Dana Farber/Harvard Cancer Center.
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Supplementary information
Supplementary Fig. 1
Reduction of fetal liver B cell progenitors in UREΔ/Δ mice. (PDF 76 kb)
Supplementary Fig. 2
Gating example of GFP+/CFP+ double transduced cells. (PDF 18 kb)
Supplementary Fig. 3
Decreasing Wnt-signaling activity during T cell maturation. (PDF 16 kb)
Supplementary Fig. 4
Lymphomas are transplantable into non-irradiated NOD/SCID recipient mice. (PDF 417 kb)
Supplementary Table 1
Genes identified by cloning lost RLGS fragments (PDF 7 kb)
Supplementary Table 2
Oligonucleotide primers and probes used for real-time RT-PCR. (PDF 8 kb)
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Rosenbauer, F., Owens, B., Yu, L. et al. Lymphoid cell growth and transformation are suppressed by a key regulatory element of the gene encoding PU.1. Nat Genet 38, 27–37 (2006). https://doi.org/10.1038/ng1679
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DOI: https://doi.org/10.1038/ng1679
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