Figure 3.

Loss of CAF-1 rescues growth of psh1Δ and cka2Δ strains and reduces Cse4 incorporation at promoters. (A) Cse4 was overexpressed from the gal promoter on a 2 micron plasmid in WT, psh1Δ, cac2Δ, cka2Δ, psh1Δ cac2Δ and cka2Δ cac2Δ strains in the W303a background. ‘EV’ indicates an empty vector control as a point of comparison. 10-fold serial dilutions of overnight cultures were plated to either SD-His or SGal-His medium. psh1Δ and cka2Δ strains overexpressing Cse4 grow poorly as expected whereas psh1Δ cac2Δ and cka2Δ cac2Δ strains grow similar to WT strains. (B) ChIP was performed for Cse4 using Cse4 overexpressing strains from (A). qPCR was used to detect Cse4 levels for the PHO5 promoter and rDNA. The y-axis indicates arbitrary units representing the enrichment for each sequence from ChIP performed with and without antibody, with respect to the signal for total chromatin for each sample. The error bars represent the standard deviation for ChIP performed in triplicate. P-values were calculated for psh1Δ cac2Δ strain relative to psh1Δ using a two-sided t-test.