Fig. 5
From: Cryo-EM structure of the human MLL1 core complex bound to the nucleosome
ASH2L Linker-IDR is important for NCP binding and methyltransferase activity. a Multiple sequence alignment of ASH2L Linker-IDR region (residues 202–254). The blue box indicated 205-KRK-207, key residues for NCP recognition. b Top, electrophoretic mobility shift assay of ASH2L and ASH2L mutants as indicated on top. Bottom, the unbound NCP in the gel image was quantified by ImageJ and presented after normalization against the NCP alone signal, which was arbitrarily set as 1 (100%). This experiment was repeated separately to confirm the main conclusions. c Immunoblot to detect in vitro histone methyltransferase activity with the NCP as the substrate. Reconstituted MLL1RWSAD complexes containing wild-type and mutant ASH2L, used as indicated on top. Immunoblots of RbBP5 and ASH2L included as controls.
