Abstract
Constitutive heterochromatin in Arabidopsis thaliana is marked by repressive chromatin modifications, including DNA methylation, histone H3 dimethylation at Lys9 (H3K9me2) and monomethylation at Lys27 (H3K27me1). The enzymes catalyzing DNA methylation and H3K9me2 have been identified; alterations in these proteins lead to reactivation of silenced heterochromatic elements. The enzymes responsible for heterochromatic H3K27me1, in contrast, remain unknown. Here we show that the divergent SET-domain proteins ARABIDOPSIS TRITHORAX-RELATED PROTEIN 5 (ATXR5) and ATXR6 have H3K27 monomethyltransferase activity, and atxr5 atxr6 double mutants have reduced H3K27me1 in vivo and show partial heterochromatin decondensation. Mutations in atxr5 and atxr6 also lead to transcriptional activation of repressed heterochromatic elements. Notably, H3K9me2 and DNA methylation are unaffected in double mutants. These results indicate that ATXR5 and ATXR6 form a new class of H3K27 methyltransferases and that H3K27me1 represents a previously uncharacterized pathway required for transcriptional repression in Arabidopsis.
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Acknowledgements
We thank C.E. Walczak for assistance with microscopy. This work was supported by grants to S.D.M. from the National Science Foundation (IOB-0447583) and National Institutes of Health (GM075060); to Y.J. from the Fonds québécois de recherche sur la nature et les technologies; and to Y.V.B. from the US Public Health Service (National Research Service award GM07104). Research in the laboratory of S.E.J. was supported by National Institutes of Health grant GM60398. S.E.J. is an investigator of the Howard Hughes Medical Institute.
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Y.J. carried out the genetic and biochemical characterization of ATXR5 and ATXR6. Y.J. and C.A.L. generated the gene expression and ChIP data. S.F. generated and sequenced the BS-Seq libraries. Y.V.B. conducted the locus-specific bisulfite sequencing analyses. H.S. validated the ChIP and RT-PCR results. C.A.L., Y.V.B. and L.M.J. conducted the immunofluorescence studies. S.C., M.P., S.F. and S.E.J. analyzed the BS-Seq data. S.D.M., M.P. and S.E.J. participated in the design of experiments.
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Jacob, Y., Feng, S., LeBlanc, C. et al. ATXR5 and ATXR6 are H3K27 monomethyltransferases required for chromatin structure and gene silencing. Nat Struct Mol Biol 16, 763–768 (2009). https://doi.org/10.1038/nsmb.1611
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DOI: https://doi.org/10.1038/nsmb.1611
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