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. 2018 Oct 16;46(21):11129–11143. doi: 10.1093/nar/gky920

Figure 4.

Figure 4.

Requirement of the Set1 N-terminal region for H3K4 methylation activity of the Set1 complex in the absence of Spp1. (A) A schematic diagram of Set1Cs containing FL Set1 and derived Set1 fragments along with associating subunits. Note that Spp1 is absent in all complexes. (B) SDS-PAGE/Coomassie blue staining of purified Set1Cs reconstituted with baculoviruses expressing FLAG-Set1 or FLAG-Set1 fragments and six (untagged) Set1C subunits. (C and D) Requirement of the Set1 N-terminal 1-229 residues for H2Bub-dependent H3K4 methylation activity of the Spp1-deficient Set1Cs. Free histone H3 (C) and H2Bub chromatin (D) were subjected to in vitro HMT assays with indicated purified Set1Cs (B). (E) Requirement of the Set1 N-terminal 1-229 resides for H3K4 methylation in yeast cells that lack Spp1. Plasmids expressing indicated HA (three copies)-tagged FL Set1 or Set1 fragments were introduced into the set1Δspp1Δ yeast strain. Yeast whole cell extracts were analyzed by immunoblotting with indicated antibodies.