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Extended Data Fig. 4 | H4K20me2 dosage dependent interaction between
ORC1 and H4K20me2 nucleosomes. a, b, Docking of the SUV420H1 and H2A.Z
nucleosome structures shows the interaction between R257 of SUV420H1 and
E64 of H4 (a), and the interaction between K333 of SUV420H1 and D97 of H2A.Z
(b). c, Western blots and^3 H autography analysis of the products of a histone
methyltransferase assay using wild-type SUV420H1 or SUV420H1 mutants.
d, Mass-spectrometry analysis of H4 histones with dimethylated KC20,
produced through chemical methylation reactions in vitro; mass spectrometry
was performed once to confirm the methylation state. e, Western blot analysis
of the interaction between H2A.Z mononucleosomes with the H4KC20me2
modification and ORC1 or ORC1 BAH-domain mutants. f, Western blots show
the interaction between ORC1 and H2A or H2A.Z mononucleosomes with
different H4KC20me2 states. g, Western blots show the interaction between
ORC1 and H2A.Z polynucleosomes with increasing ratios of a H4KC20me2
octamer. The western blots in panels c, e–g and the^3 H autography in panel c
were independently repeated twice with similar results. H4 was used as a
loading control in panel c. H3 was used as a loading control in panels e–g. For
gel source data, see Supplementary Fig. 1.