FIG. 2.

Yju2 is required for pre-mRNA splicing in vivo. (A) Protein sequence alignment of Yju2 orthologs using MAFFT and GeneDoc. Putative Yju2 orthologs were identified by using published KEGG (Kyoto Encyclopedia of Genes and Genomes) databases (24). Conserved residues are shaded black for 100% conservation, gray for 80% conservation, and light gray for 60% conservation. Abbreviations, protein accession numbers (from NCBI), and percentages of identity with Yju2 are the following: Sc, Saccharomyces cerevisiae, NP_012828, 100%; Sp, Schizosaccharomyces pombe, NP_001018234, 40%; Hs, Homo sapiens, NP_060544, 36.32%; Rn, Rattus norvegicus, XP_576700, 36.32%; Mm, Mus musculus, NP_082657, 36.32%; Dm, Drosophila melano-gaster, NP_611092, 35.38%; At, Arabidopsis thaliana, NP_173156, 33.99%; and Ce, Caenorhabditis elegans, NP_498576, 33.33%. (B) Growth curves of GAL-YJU2 cells in glucose medium (YPD) and galactose medium (YPG). Cells were grown in galactose medium to mid-log phase and then either continuously maintained in YPG or shifted to YPD. Cells were collected at 0, 4, 8, 12, 16, 20, 24, and 28 h after the shift for measurements of the optical density at 600 nm (OD₆₀₀). (C) Protein extracted from collected cells was analyzed by probing Western blots with the anti-Yju2 antibody. Anti-G6PDH antibody was used as an internal control. (D) Total RNA extracted from collected cells was analyzed by primer extension using R13 as the primer. The prp2 mutant was grown at 37°C for 2 h before harvest. YPD, yeast extract-peptone-dextrose; YPG, yeast extract-peptone-galactose; Gal, galactose; Glc, glucose.