Abstract
Histone H3 Lys4 (H3K4) methylation is a prevalent mark associated with transcription activation. A common feature of several H3K4 methyltransferase complexes is the presence of three structural components (RbBP5, Ash2L and WDR5) and a catalytic subunit containing a SET domain. Here we report the first biochemical reconstitution of a functional four-component mixed-lineage leukemia protein-1 (MLL1) core complex. This reconstitution, combined with in vivo assays, allows direct analysis of the contribution of each component to MLL1 enzymatic activity and their roles in transcriptional regulation. Moreover, taking clues from a crystal structure analysis, we demonstrate that WDR5 mediates interactions of the MLL1 catalytic unit both with the common structural platform and with the histone substrate. Mechanistic insights gained from this study can be generalized to the whole family of SET1-like histone methyltransferases in mammals.
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Acknowledgements
We are grateful to J. Wysocka (Rockefeller University) and W. Herr (University of Lausanne) for anti-WDR5, Z. Fu in the Roeder laboratory for technical support and Y. Liu in the Allis laboratory for critical reading of the manuscript. Y.D. is a fellow of the Irvington Institute for Immunological Research and T.A.M. is a fellow of the Canadian Institutes of Health Research. This work was supported by US National Institutes of Health grants (to R.G.R., J.W.L., G.L.V. and C.D.A.) and by funds from the Rockefeller University (to R.G.R.).
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Y.D. was responsible for reconstitution of the MLL1 core complex, in vitro enzymatic assays and RNA interference experiments, T.A.M. was responsible for ChIP analysis and RT-PCR after siRNA treatment, S.H.L. mapped the RbBP5-Ash2L interaction interface and A.J.R. pinpointed the WDR5 mutations that disrupt substrate binding. J.W.L., G.L.V., C.D.A. and R.G.R. supervised the project and take overall responsibility for their joint research. All authors discussed the results and commented on the manuscript.
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Dou, Y., Milne, T., Ruthenburg, A. et al. Regulation of MLL1 H3K4 methyltransferase activity by its core components. Nat Struct Mol Biol 13, 713–719 (2006). https://doi.org/10.1038/nsmb1128
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DOI: https://doi.org/10.1038/nsmb1128
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