Fig. 1

A methyl-donor analogue-based screen identified ATF7IP as a G9a/GLP substrate. a A scheme of ProSeAM-based screen of substrates for G9a or GLP methylation. L and H indicate light-(¹²C6) and heavy-(¹³C6) lysine labelling, respectively. b Western blot analysis with streptavidin-HRP shows a labelling of biotin tag to endogenous proteins in the cell lysates. Blot with anti-H3 antibody was used as a loading control. c A Venn diagram of identified proteins in the screen. Shared candidates were ATF7IP, LIG1, and ARFG2. Identified histone H3K9-like sequences within substrates were shown. Validation experiments were done in this study (ATF7IP and Nup107) or previous studies (Wiz and LIG1; [16, 19]). d A schematic of human and mouse ATF7IP protein. Known regions that interact with other proteins were indicated in colour. A histone H3K9-like sequence locates in the N-terminus of ATF7IP. The histone H3K9-like motif is conserved among vertebrates. e An in vitro methylation assay with recombinant ATF7IP protein (residues 1–364), GST-tagged SET-domain of G9a or GLP, and ¹⁴C-labelled SAM. An incorporation of the methyl-moiety was detected as an autoradiographic signal. WT and KR indicate wild-type and K16R mutant of ATF7IP, respectively