A structural “toggle” in the RNaseH domain of Prp8 helps balance splicing fidelity and catalytic efficiency. Saccharomyces cerevisiae

Purpose: To assess the global impacts of RNaseH domain alleles of Prp8 on the in vivo splice site selection for native introns. Conclusions: Our study reveals functional links between distinct spliceosomal conformations and splicing fidelity. Overall design: Lariat sequencing of PRP8 WT and four mutants in the RNaseH domain of Prp8 in the background of Δdbr1 to profile global locations of splice site activation and compare aberrant activation rate across the five strains. Two additional lariat sequencing libraries were prepared as technical replicates for a second biological sample of strain prp8-N1869D to assess the technical reproducibility of the method.