FIG. 2.
(A) SpMst1p and SpMst2p are nuclear- and chromatin-bound throughout the cell cycle. The wild type (FY258) and strains expressing Mst1-HA (FY1812), Mst2-HA (FY1818), Mcm2-HA (FY798), and Orp1-HA (FY793) were permeabilized and washed with the indicated concentrations of Triton X-100. DNA was visualized by DAPI staining (DNA) and HA-tagged proteins using the HA antibody and a secondary antibody conjugated to Cy3 (HA). Bar, 10 μm. (B to E) Δmst2 cells are sensitive to HU (A) and slightly sensitive to MMS (D) and UV irradiation (E). For HU, TBZ, and MMS sensitivity analyses, cells were grown in liquid cultures to an A595 of 1, serially diluted fivefold, spotted on the indicated plates, and incubated at 25°C for 3 to 5 days. Equal numbers of cells were spotted on each plate. To analyze UV sensitivity, 1,000 cells were plated on YES, irradiated with 50, 100, 150, 200, or 300 J/m2, and incubated at 23°C for 3 days in the dark. Cells were counted and the percentage survival with respect to the number of colonies on the untreated plates was plotted versus the UV dose. Strains used include the wild type (FY258), Δrad3 (FY1107), hsk1ts (FY2104), Δmst2 (FY1889), rad21ts (FY1159), Δmst2 rad21ts (FY2032), Δswi6 (FY2056), Δtaz1 (FY1421), and Δmst2 Δtaz1 (FY2330 and FY2331). WT, wild type.