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A two-state activation mechanism controls the histone methyltransferase Suv39h1

Nature Chemical Biology volume 12, pages 188–193 (2016)Cite this article

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Abstract

Specialized chromatin domains contribute to nuclear organization and regulation of gene expression. Gene-poor regions are di- and trimethylated at lysine 9 of histone H3 (H3K9me2 and H3K9me3) by the histone methyltransferase Suv39h1. This enzyme harnesses a positive feedback loop to spread H3K9me2 and H3K9me3 over extended heterochromatic regions. However, little is known about how feedback loops operate on complex biopolymers such as chromatin, in part because of the difficulty in obtaining suitable substrates. Here we describe the synthesis of multidomain 'designer chromatin' templates and their application to dissecting the regulation of human Suv39h1. We uncovered a two-step activation switch where H3K9me3 recognition and subsequent anchoring of the enzyme to chromatin allosterically promotes methylation activity and confirmed that this mechanism contributes to chromatin recognition in cells. We propose that this mechanism serves as a paradigm in chromatin biochemistry, as it enables highly dynamic sampling of chromatin state combined with targeted modification of desired genomic regions.

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Figure 1: Heterotypic designer chromatin substrates.
Figure 2: Reconstitution of Suv39h1-dependent heterochromatin spreading in vitro.
Figure 3: Trans activation of Suv39h1.
Figure 4: The N terminus of Suv39h1 contributes to chromatin binding in vitro.
Figure 5: The Suv39h1 N terminus contributes to chromatin binding in vivo.

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Acknowledgements

We thank U. Nguyen and Y. David for help with tissue culture; G. Laevsky for advice on microscopy; P. Lewis for providing biotinylated H3 peptides; C.D. Allis, G. Debelouchina, Z. Brown, B. Wang and C. Jenness for helpful discussions; and K. Jani for careful proofreading of this manuscript. NIH-3T3 cells were a gift from J. Schwarzbauer (Princeton University). Funding provided by the Swiss National Science Foundation (postdoctoral fellowships to M.M.M. and B.F.) and the US National Institutes of Health (grant R01-GM107047 to T.W.M.).

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  1. Beat Fierz

    Present address: Present address: Ecole Polytechnique Fédérale de Lausanne, Lausanne, Switzerland.,

Authors and Affiliations

  1. Department of Chemistry, Princeton University, Princeton, New Jersey, USA

    Manuel M Müller, Beat Fierz, Lenka Bittova, Glen Liszczak & Tom W Muir

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Contributions

M.M.M., B.F. and T.W.M. conceived the project; M.M.M., B.F., L.B. and G.L. designed and performed experiments with supervision from T.W.M.; all authors analyzed data; M.M.M. and T.W.M. wrote the manuscript with contributions from all authors.

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Correspondence to Tom W Muir.

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Müller, M., Fierz, B., Bittova, L. et al. A two-state activation mechanism controls the histone methyltransferase Suv39h1. Nat Chem Biol 12, 188–193 (2016). https://doi.org/10.1038/nchembio.2008

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