FIGURE 3.

Defects of dna2-K1080E can be corrected by active Rad27. A, the Rad52-dependent suppression of dna2-K1080E was examined in rad50-null cells. Overexpression of Rad52 in rad50-null cells is indicated as rad50Δ+Rad52. Cells were grown and spotted as described in the legend for Fig. 1A. FOA, 5-fluorooritic acid. B, the YJA1BJK65 cells were transformed with pRS325-ADH1 plasmids expressing Rad27, Rad27-FLAG (Rad27 with a FLAG tag fused to its C terminus), Rad27-DA (Rad27-D179A; catalytically defective), and Rad27-DA-FLAG. Transformants were grown and spotted as described above in the legend for Fig. 1A. C, expression levels of Rad27-FLAG and Rad27-DA-FLAG were examined by Western blot using anti-FLAG antibody (α-FLAG; F-3165, Sigma-Aldrich). Histone H3 was used as the loading control. α-H3, antibodies against histone H3. The experiment was done as described in the legend for Fig. 1B.