Whole-genome splicing profile of a fission yeast strain expressing Prp4_as2 kinase.

<p>(A) The frequency histograms assign the number of introns found for each calculated Relative Splicing Efficiency Index (RSEI) in the absence (- Inhibitor) of 1NM-PP1 or after 30 and 60 min in the presence of inhibitor. The bin size is 0.05. Bars with negative RSEI values display Prp4-dependent introns (1008 introns) while bars with positive RSEI values represent Prp4-independent introns (2557 introns). The dashed line marks the 0 value. (B) Similar size distributions of Prp4-independent and -dependent introns. (C) A hypothetical example of a fission yeast pre-mRNA. Consensus sequences of the 5’ SS, branch sequence with branch point A (bp, arrow), and 3’ SS are shown. These consensus sequences do not differ between Prp4-dependent and–independent introns. The sequence logos were generated using WebLogo [<a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1005768#pgen.1005768.ref074" target="_blank">74</a>]. (D) Splicing of the introns of <i>rpb5</i>, <i>tbp1</i>, and <i>mrp17</i> monitored by RT-PCR using specific primers, as indicated in the schemes above the images, in the absence of inhibitor (-Inh) or after 10 and 30 minutes in the presence of inhibitor (+Inh). Asterisks indicate the expected position of fragments if the introns are not spliced out. RSEI below the images was obtained from cells collected after 30 min in the presence of inhibitor. Roman numerals indicate the 5’→3’ order of introns. The numbers on the left side of the image represent the sizes of the DNA fragments (bp). M, DNA size marker.