Abstract
We have developed a readily accessible disulfide-directed methodology for the site-specific modification of histones by ubiquitin and ubiquitin-like proteins. The disulfide-linked analog of mono-ubiquitylated H2B stimulated the H3K79 methyltransferase activity of hDot1L to a similar extent as the native isopeptide linkage. This permitted structure-activity studies of ubiquitylated mononucleosomes that revealed plasticity in the mechanism of hDot1L stimulation and identified surfaces of ubiquitin important for activation.
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Acknowledgements
We acknowledge J. Fernandez, L. Miller and R. Angeletti for assistance with ESI-MS analysis. We thank J. Kim and R. Subramanian for help in preparing hDot1 and M. Pratt for stimulating discussions. This work was funded by the US National Institutes of Health (grant number RC2CA148354) and the Starr Cancer Consortium. B.F. was supported by the Novartis and Swiss National Science Foundations. R.K.M. was supported by a US National Institutes of Health Medical Scientist Training Program grant.
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C.C. and T.W.M. designed the experiments. C.C. performed the experiments. C.C., R.K.M. and B.F. prepared new reagents. C.C. and T.W.M. analyzed the experimental results and wrote the manuscript.
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Chatterjee, C., McGinty, R., Fierz, B. et al. Disulfide-directed histone ubiquitylation reveals plasticity in hDot1L activation. Nat Chem Biol 6, 267–269 (2010). https://doi.org/10.1038/nchembio.315
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DOI: https://doi.org/10.1038/nchembio.315
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