FIGURE 4.

Detection of the loss of 2′O-methylation in yeast rRNA. (A) Ligation reactions carried out using yeast total RNA from the wild-type or snoRNA mutant strains using D- or N-oligos. “Standard” corresponds to the product derived from a known amount of the model 30-mer RNA added to the ligation mixture. ΔsnR53 corresponds to a loss of 2′O-methylation at A796 in 18S rRNA, and ΔsnR67 corresponds to a loss of 2′O-methylation at G2618 in 25S rRNA. In this panel, the ligation reaction was performed for 120 min. (B) Relative amount of ligation product at the A796 or G2618 site for the snoRNA mutant strains over the wild-type strain. The decreased ratio at the latest time (120 min) is due to increased amount of product in the wild-type strain, as the amount of product in the mutant strains approaches saturation.