Figure 5.

Set1 catalytic activity but not its recruitment to chromatin is required for its gene protective function. (A) Diagram of Set1 constructs including WT-Set1 and two point mutations in the catalytic SET domain: C862A and G852S. Constructs are expressed from the set1 promoter at the native set1 locus with an N-terminal 2xFlag tag. (B) Licor Western blot for H3K4me3 with GAPDH loading control of whole-cell extracts from wild-type untagged Set1, the Δset1 parent of the 2xFlag constructs, 2xFlag-wtSet1, 2xFlag-Set1C862A, and 2xFlag-Set1G852S. (C) Histogram plots as in Figure 1 of normalized “orange” signal from rpl401p:HSS Δboundary isolates that were transformed with either untagged wt-Set1, Set1C862A, or Set1G852S. (D) Anti-Flag ChIP-qPCR data in genetic backgrounds as in B. Error bars represent 1SD from two technical replicate ChIPs.