Recombinant Mononucleosomes H3.1 (K18I)

In vivo, histones are wrapped around by DNA in chromatin. Therefore, nucleosomes are more physiologically relevant substrates than histones and histone-derived peptides for in vitro studies. More importantly, some histone methyltransferases are significantly more active, as well as specific, when using nucleosomal substrates in HMT assays, such as DOT1L and NSD family enzymes. Nucleosomes are also widely used in histone methyltransferase screening assays to identify small molecular inhibitors for drug discovery. Histones are linked to tumorigenesis primarily through alterations in their PTMs and the enzymes regulating these modifications, suggesting that they might disrupt the reading, writing, and/or erasing of these marks. Mutations in histone H3 occur with high genetic penetrance within rare pediatric gliomas and sarcomas. In H3 variants, the mutation is most often a lysine-to-methionine (K-M) mutation, occasionally glycine mutations (G34R/V/W/L) occur too. More K-to-M/I mutations were observed, raising the possibility that the functional effects associated with known K-to-M/I changes (that is, function in a dominant fashion to block the methylation of corresponding lysines on wild type histones) may extend to additional contexts. Researchers found that mutations in the subset with a TMB ? 2 mutations per Mb included H3 (K27M) and H3 (G34W), and other mutations like H3 (E105K/Q), mutations at H3 N-terminal residues at or near PTM sites including R2, R8, K18 and R26, as well as residues in the acidic patch such as H2A residues E56, E64, E9, and E92 and H2B residues E105 and E133, which might act as oncohistones. According to the structural analysis, specifically for the KDM6 family, the hydrophobic mutation - histone H3 K18I- showed a significantly higher affinity towards the KDM6 enzymes.