FIGURE 5.

C-terminal PHDs of Msc1 are important for function and for association with Swr1. A, shown is schematic representation of the deletion mutants of Msc1 carried by the pSP1 plasmid. All constructs also had a 3×HA tag at the C terminus. B, the indicated plasmids were introduced into the msc1Δ mis6-302 (NW1703; left panels) and msc1Δ mis12-537 (NW1617; right panels) strains and grown at 25 °C to mid-log phase. Serial dilutions were spotted on YEA plates and incubated at the indicated temperatures for 3 days. C, a strain with an integrated FLAG-tagged allele of swr1 and deletion of the msc1 gene (NW2646) was transformed with the indicated plasmids. Cells were grown to mid-log phase, lysed, and subjected to immunoprecipitation with antibody to FLAG-tagged Swr1. Precipitates were separated by SDS-PAGE, transferred to nitrocellulose, and incubated with antibodies to detect Swr1-FLAG (upper panel) and Msc1-HA (lower panel).