Epigenetic histone modification: systematic regulation of transcription
1. In vitro assay system of histone modifications
To measure the catalytic activities of histone modifiers, we are highly skilled at recombinant protein purification and have numerous types of enzyme constructs. Using core histones and nucleosomes, we identified for the first time, RE-IIBP is a histone H3K79 methyltransferase and KDM2B as a H3K79 demethylase.
Detection of H3K79 methylation by RE-IIBP and demethylation by KDM2B via in vitro assay and mass spectrometry
2. In vivo analysis of histone modification patterns
Histones or nucleosomes extracted from cells are analyzed using specific antibodies for each histone PTM to evaluate global patterns. By mapping global histone PTMs sites, we identified that which regions are affected by certain histone modifiers and whether those alterations are specific for target genes or global regulation. ChIP-seq using H3K79me3 antibody has shown that depletion of KDM2B leads to genome-wide accumulation of H3K79 methyl marks.
Analysis of effects of KDM2B on H3K79me3 level in the whole genome