Shu1 localizes to the nucleolus and affects rDNA recombination. (A) Yeast with Shu1-YFP-YFP and Nop1-CFP were analyzed by fluorescence microscopy for colocalization (Merge). (B) The frequency of rDNA recombinants (CANR, ade2) was measured in WT, shu1Δ, uaf30Δ, and uaf30Δ shu1Δ yeast strains, and they were plotted with SD. (C) The amounts of rDNA in WT, shu1Δ, uaf30Δ, and uaf30Δ shu1Δ strains were quantitated by analyzing the amount of rDNA resulting from restriction digest of total DNA, revealing a 9.1-kb fragment as described in Materials and Methods. SD are plotted. (D) WT and uaf30Δ strains were transformed with the empty vector (pWJ1530) or the SHU1 overexpression plasmid (pWJ1530-SHU1). Strains were grown in the presence of 100 μM copper (CuSO4), and the frequency of rDNA recombinants (CANR, ade2) was measured and the SE are plotted. Note that the recombination frequencies in uaf30Δ cells were different relative to (B), likely due to their growth in synthetic minimal medium, which was needed to maintain the plasmid.
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