title: Interconnections Between RNA-Processing Pathways Revealed by a Sequencing-Based Genetic Screen for Pre-mRNA Splicing Mutants in Fission Yeast.
abstract: Pre-mRNA splicing is an essential component of eukaryotic gene expression and is highly conserved from unicellular yeasts to humans. Here, we present the development and implementation of a sequencing-based reverse genetic screen designed to identify nonessential genes that impact pre-mRNA splicing in the fission yeast Schizosaccharomyces pombe, an organism that shares many of the complex features of splicing in higher eukaryotes. Using a custom-designed barcoding scheme, we simultaneously queried ∼3000 mutant strains for their impact on the splicing efficiency of two endogenous pre-mRNAs. A total of 61 nonessential genes were identified whose deletions resulted in defects in pre-mRNA splicing; enriched among these were factors encoding known or predicted components of the spliceosome. Included among the candidates identified here are genes with well-characterized roles in other RNA-processing pathways, including heterochromatic silencing and 3' end processing. Splicing-sensitive microarrays confirm broad splicing defects for many of these factors, revealing novel functional connections between these pathways.
authors.affiliation: Department of Molecular Biology and Genetics, Cornell University, Ithaca, New York 14853.
authors.forename: Amy
authors.initials: A
authors.lastname: Larson
authors.affiliation: Department of Molecular Biology and Genetics, Cornell University, Ithaca, New York 14853.
authors.forename: Benjamin Jung
authors.initials: BJ
authors.lastname: Fair
authors.affiliation: Department of Molecular Biology and Genetics, Cornell University, Ithaca, New York 14853 jpleiss@cornell.edu.
authors.forename: Jeffrey A
authors.initials: JA
authors.lastname: Pleiss
mesh_terms: D005820:Genetic Testing; D055106:Genome-Wide Association Study; D006570:Heterochromatin / Q000235:genetics / Q000378:metabolism; D008954:Models, Biological; D009154:Mutation; D012322:RNA Precursors / Q000235:genetics*; D012323:RNA Processing, Post-Transcriptional; D012326:RNA Splicing; D000072260:RNA Splicing Factors / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics; D012568:Schizosaccharomyces / Q000235:genetics* / Q000378:metabolism; D017422:Sequence Analysis, DNA
chemical_list: D006570:Heterochromatin; D012322:RNA Precursors; D000072260:RNA Splicing Factors; D012333:RNA, Messenger
keywords: Schizosaccharomyces pombe; genetic screen; genomics; heterochromatin; pre-mRNA splicing