Abstract
Transcription factors are frequently altered in leukaemia through chromosomal translocation, mutation or aberrant expression1. AML1–ETO, a fusion protein generated by the t(8;21) translocation in acute myeloid leukaemia, is a transcription factor implicated in both gene repression and activation2. AML1–ETO oligomerization, mediated by the NHR2 domain, is critical for leukaemogenesis3,4,5,6, making it important to identify co-regulatory factors that ‘read’ the NHR2 oligomerization and contribute to leukaemogenesis4. Here we show that, in human leukaemic cells, AML1–ETO resides in and functions through a stable AML1–ETO-containing transcription factor complex (AETFC) that contains several haematopoietic transcription (co)factors. These AETFC components stabilize the complex through multivalent interactions, provide multiple DNA-binding domains for diverse target genes, co-localize genome wide, cooperatively regulate gene expression, and contribute to leukaemogenesis. Within the AETFC complex, AML1–ETO oligomerization is required for a specific interaction between the oligomerized NHR2 domain and a novel NHR2-binding (N2B) motif in E proteins. Crystallographic analysis of the NHR2–N2B complex reveals a unique interaction pattern in which an N2B peptide makes direct contact with side chains of two NHR2 domains as a dimer, providing a novel model of how dimeric/oligomeric transcription factors create a new protein-binding interface through dimerization/oligomerization. Intriguingly, disruption of this interaction by point mutations abrogates AML1–ETO-induced haematopoietic stem/progenitor cell self-renewal and leukaemogenesis. These results reveal new mechanisms of action of AML1–ETO, and provide a potential therapeutic target in t(8;21)-positive acute myeloid leukaemia.
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Acknowledgements
We thank N. A. Speck and J. H. Bushweller for providing the AML1–ETO m7 plasmid, and R. Baer for providing anti-SCL antibodies. This work was supported by National Institutes of Health (NIH) grants CA163086 (R.G.R.), CA129325 (R.G.R.), CA113872 (R.G.R.) and CA166835 (S.D.N.), Starr Cancer Consortium grant I5-A554 (R.G.R., D.J.P. and S.D.N.), Leukemia and Lymphoma Society (LLS) SCOR grants 7013-02 (R.G.R. and S.D.N.) and 7132-08 (R.G.R., A.M. and D.J.P.), and Rockefeller University Center for Clinical and Translational Science Pilot Project grant UL1RR024143 from NIH (X.-J.S.). X.-J.S. was a Starr Cancer Consortium Visiting Fellow. L.W. was an Empire State Stem Cell Scholar and an LLS Fellow. Y.J. was an American Society of Haematology Scholar. W.-Y.C. was an LLS Fellow. D.J.P. was supported by funds from the Abby Rockefeller Mauze Trust and the Maloris Foundation.
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X.-J.S. and R.G.R. conceived the project. R.G.R. supervised the biochemical studies. S.D.N. supervised the leukaemia pathological studies. D.J.P. supervised the structural studies. A.M. supervised the genomic studies. X.-J.S., Z.W., L.W., Y.J., N.K., T.D.S., W.-Y.C., Z.T., T.N., O.E. and W.F. performed the experiments and analysed the data. X.J.S. and R.G.R. wrote the paper.
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This file contains Supplementary Figures 1-15, Supplementary Tables 1-3, a Supplementary Discussion and additional references. (PDF 2895 kb)
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Sun, XJ., Wang, Z., Wang, L. et al. A stable transcription factor complex nucleated by oligomeric AML1–ETO controls leukaemogenesis. Nature 500, 93–97 (2013). https://doi.org/10.1038/nature12287
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DOI: https://doi.org/10.1038/nature12287
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