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. 2017 Dec;23(12):1834–1849. doi: 10.1261/rna.062794.117

FIGURE 2.

FIGURE 2.

Pseudouridines in Toxoplasma snRNAs are homologous to those in Saccharomyces cerevisiae and human snRNAs, and are not TgPUS1-dependent. (A) Primer extension of Toxoplasma U6 snRNA using a primer targeting the 3′-end of the U6 transcript. RNA from uninfected HFF cultures (UI) and HFF cultures infected with wild-type (WT) or TgPUS1-deficient (Mut) tachyzoites and bradyzoites was analyzed following treatment with CMCT (+) or mock treatment (−), as indicated. The 3′-end of the primer is located 25 residues from the putative 3′-end of the U6 snRNA. Radiolabeled cDNAs were resolved on an 8% polyacrylamide gel. Unextended primer is seen at the bottom of the gel (black arrowhead) while the location of the putative Ψ is indicated with an arrow. The migration of size markers (given in nucleotides) are indicated on the left. (B) As in A, but showing primer extension of Toxoplasma U2 snRNA. The 3′-end of the primer is located 87 residues from the putative 3′-end of the U2 snRNA.