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Custom Vector Construction> | MATERIALS AND METHODS Protein expression and purification Open reading frames (ORFs), optimized for overexpression in Escherichia coli, of Ricinus communis and Glycine max ATXR5 (RcATXR5 and GmATXR5 respectively) were purchased from GenScript and subcloned in the parallel expression vector pGST2.... 1–10, ARTKQTARKSY) encompassing K4, were synthesized by Genscript and solubilized in 100 mM Tris (pH 8. | Get A Quote |
In plants, the histone H3.1 lysine 27 (H3K27) mono-methyltransferases ARABIDOPSIS TRITHORAX RELATED PROTEIN 5 and 6 (ATXR5/6) regulate heterochromatic DNA replication and genome stability. Our initial studies showed that ATXR5/6 discriminate between histone H3 variants and preferentially methylate K27 on H3.1. In this study, we report three regulatory mechanisms contributing to the specificity of ATXR5/6. First, we show that ATXR5 preferentially methylates the R/F-K*-S/C-G/A-P/C motif with striking preference for hydrophobic and aromatic residues in positions flanking this core of five amino acids. Second, we demonstrate that post-transcriptional modifications of residues neighboring K27 that are typically asso... More