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. 2011 Jan 14;286(10):8369–8374. doi: 10.1074/jbc.M110.203380

FIGURE 3.

FIGURE 3.

The hASH1L catalytic domain methylates histone H3 Lys-36. A, HMTase assay of the hASH1L catalytic domain was performed with wild-type hASH1L (WT), F2260A, and the hyperactive mutant (Q2265A) (lanes 1, 2, and 3) using recombinant mononucleosomes. Recombinant mononucleosomes (Nuc) and HeLa nucleosomes (Hela Nuc) were used as controls for the antibodies (lanes 4 and 5). The methylation product of the hASH1L catalytic domain is specifically recognized by the anti methyl-H3K36 antibody but not by the methyl-H3K4 antibody. The hyperactive mutant (Q2265A) is able to also produce H3K36 trimethylation. B and C, the wild-type hASH1L SET catalytic domain and the hyperactive mutant methylate the wild-type or H3K4A mutant histone, octamer, and nucleosome (Nuc.), but not the H3K36A mutant histone, octamer, and nucleosome.