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. 2018 Jan;24(1):30–42. doi: 10.1261/rna.063842.117

FIGURE 6.

FIGURE 6.

Prior pseudouridylation of U2 snRNA at position 43 inhibits positioning of Ψ44 by mouse SCARNA8 and S. pombe pugU2-43/44, SPNCRNA.1709. Fluorescent primer extension reactions. (A) Mouse SCARNA8 was expressed in the S. cerevisiae strains pus1Δ (top magenta trace) and pus7Δ (bottom green trace) from a plasmid with a weak CYC1 promoter that allowed prior modification of position 44 in yeast U2 snRNA by endogenous Pus1p. In the pus7Δ strain, modification of position 45 was reduced relative to that in the pus1Δ strain (arrow shows reduction in peak height). Stars indicate SCARNA8-induced pseudouridylation. Modification of position 35 in the pus7Δ strain served as an internal positive control. (B) Top trace (red): Pseudouridylation pattern of U2 snRNA in wild-type S. pombe (strain ED666). Bottom trace (blue): Overexpression of pugU2-43/44ΔUmut RNA caused reduction of pseudouridylation at position 44 (arrow shows reduction in peak height).