Metabolic control of histone demethylation

To analyze whether metabolites regulate histone methylation in plants, we examined genome-wide histone 3 lysine-27 trimethylation (H3K27me3) in an Arabidopsis dehydrogenase mutant (ogdh1/2) by ChIP-seq. We found that the levels of H3K27me3 were reduced at various loci in ogdh1/2 relative to wild type, indicating that metabolites control histone demethylation in Arabidopsis. Nuclei were extracted from 10-d-old WT (Col-0) and ogdh (double mutant) seedlings as previously described (Louwers et al., 2009), followed by chromatin immunoprecipitation with Magna ChIP A/G kit (Millipore). DNA libraries were constructed using NEBNext Ultra II DNA Library Prep Kit for Illumina (NEB), and subsequently were sequenced on NovaSeq 6000 (Illumina). Three biologically independent samples have been analyzed.