Figure 3.

Mutation of a conserved residue of Bcd1 hampers the ability of the protein to interact with RNA. A, strep-tagged Bcd1 (WT) or Bcd1-D72A was used to pull down snoRNPs from yeast cells. Protein and RNA from input and eluates were analyzed by Western and Northern blotting. B, quantification of the eluted RNA relative to input is shown in A. Data from two independent experiments were averaged. Error bars indicate mean ± S.E. C, EMSA shows that Bcd1 binds to snR51 (a box C/D snoRNA), albeit weakly, whereas Bcd1-D72A binding to the snoRNA is weaker. D, quantification of the data shown in C. Data show averages from four independent experiments. Errors bars indicate mean ± S.E.