Abstract
The Arabidopsis KRYPTONITE gene encodes a member of the Su(var)3-9 family of histone methyltransferases. Mutations of kryptonite cause a reduction of methylated histone H3 lysine 9, a loss of DNA methylation, and reduced gene silencing. Lysine residues of histones can be either monomethylated, dimethylated or trimethylated and recent evidence suggests that different methylation states are found in different chromatin domains. Here we show that bulk Arabidopsis histones contain high levels of monomethylated and dimethylated, but not trimethylated histone H3 lysine 9. Using both immunostaining of nuclei and chromatin immunoprecipitation assays, we show that monomethyl and dimethyl histone H3 lysine 9 are concentrated in heterochromatin. In kryptonite mutants, dimethyl histone H3 lysine 9 is nearly completely lost, but monomethyl histone H3 lysine 9 levels are only slightly reduced. Recombinant KRYPTONITE can add one or two, but not three, methyl groups to the lysine 9 position of histone H3. Further, we identify a KRYPTONITE-related protein, SUVH6, which displays histone H3 lysine 9 methylation activity with a spectrum similar to that of KRYPTONITE. Our results suggest that multiple Su(var)3-9 family members are active in Arabidopsis and that dimethylation of histone H3 lysine 9 is the critical mark for gene silencing and DNA methylation.
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Acknowledgments
We thank L. Cahoon, E. Huang, J. Bruder, and C. Hyun for technical assistance, A. Meister for flow-sorting of nuclei, E. Selker and H. Tamaru for providing the GST-DIM5 fusion construct, and members of the Jacobsen laboratory for discussions and critical review of the manuscript. This work was supported by NIH grant GM60398 (S.E.J.), NIH training grant GM07104 (J.P.J.), grants from the Land Sachsen-Anhalt (3233A/0020L) and DFG (Schu 951/8-2) (Z.J. and I.S.), and NIH grants GM49245 and GM61355 (X.Z. and X.C.).
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Jackson, J.P., Johnson, L., Jasencakova, Z. et al. Dimethylation of histone H3 lysine 9 is a critical mark for DNA methylation and gene silencing in Arabidopsis thaliana . Chromosoma 112, 308–315 (2004). https://doi.org/10.1007/s00412-004-0275-7
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DOI: https://doi.org/10.1007/s00412-004-0275-7