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A stable transcription factor complex nucleated by oligomeric AML1–ETO controls leukaemogenesis

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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Figure 1: AML1–ETO resides in and functions through AETFC.
Figure 2: The oligomerized AML1–ETO NHR2 domain mediates a specific interaction with E proteins.
Figure 3: Structural details of the NHR2–N2B interaction.
Figure 4: Role of the AML1–ETO–E-protein interactions in human HSPC self-renewal and mouse leukaemogenesis.

Accession codes

Accessions

Gene Expression Omnibus

Protein Data Bank

Data deposits

ChIP-seq and RNA-seq data have been deposited in the Gene Expression Omnibus under accession GSE43834. The crystal structure of the NHR2–N2B complex has been deposited in the Protein Data Bank under accession 4JOL.

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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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Authors

Contributions

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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Correspondence to Robert G. Roeder.

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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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