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Proteins, Expression, Isolation and Analysis> | The supernatant was incubated with Flag–agarose beads (Genscript G1 a-DYKDDDDK, 0.5 µl beads per ml culture) for 3 h at 4°C. Following incubation, the beads were washed three times with TGN (25 mM Tris, 10% glycerol, 0.02% IGEPAL CA 630, pH 7.5) plus 350 mM NaCl, and a final wash with TGN plus 120 mM NaCl. | Get A Quote |
Chemical modifications of histones can mediate diverse DNA-templated processes, including gene transcription. Here we provide evidence for a class of histone post-translational modification, serotonylation of glutamine, which occurs at position 5 (Q5ser) on histone H3 in organisms that produce serotonin (also known as 5-hydroxytryptamine (5-HT)). We demonstrate that tissue transglutaminase 2 can serotonylate histone H3 tri-methylated lysine 4 (H3K4me3)-marked nucleosomes, resulting in the presence of combinatorial H3K4me3Q5ser in vivo. H3K4me3Q5ser displays a ubiquitous pattern of tissue expression in mammals, with enrichment observed in brain and gut, two organ systems responsible for the bulk of 5... More