GSE68022
Identification of New Branch Points and Unconventional Introns in Saccharomyces cerevisiae
Spliced messages constitute one-fourth of  expressed mRNAs in the yeast Saccharomyces cerevisiae, and most mRNAs in metazoans. Splicing requires 5'' splice site (5''SS), branch point (BP), and 3'' splice site (3''SS) elements, but the role of the BP in splicing control is poorly understood because BP identification remains difficult.  We developed a high-throughput method, Branch-seq, to map BP and 5''SS of isolated RNA lariats. Applied to S. cerevisiae, Branch-seq detected 76% of expressed, annotated BPs and identified a comparable number of novel BPs.  We used RNA-seq to confirm associated 3''SS locations, identifying some 200 novel splice junctions, including an AT-AC intron.  We show that several yeast introns use two or even three different BPs, with effects on 3''SS choice, protein coding potential, or RNA stability and identify novel introns whose splicing changes during meiosis or in response to stress. Together, these findings reveal BP-based regulation and demonstrate unanticipated complexity of splicing in yeast. Overall design: 1 Lariat-seq experiment library. 3 barcoded Branch-seq libraries that make up one experiment. 26 RNA-seq samples, 2 biological replicates of each.