title: Rapid identification of mRNA processing defects with a novel single-cell yeast reporter.
abstract: It has become increasingly evident that gene expression processes in eukaryotes involve communication and coordination between many complex, independent macromolecular machines. To query these processes and to explore the potential relationships between them in the budding yeast Saccharomyces cerevisiae, we designed a versatile reporter using multicolor high-throughput flow cytometry. Due to its design, this single reporter exhibits a distinctive signature for many defects in gene expression including transcription, histone modification, pre-mRNA splicing, mRNA export, nonsense-mediated decay, and mRNA degradation. Analysis of the reporter in 4967 nonessential yeast genes revealed striking phenotypic overlaps between chromatin remodeling, histone modification, and pre-mRNA splicing. Additionally, we developed a copper-inducible reporter, with which we demonstrate that 5-fluorouracil mimics the mRNA decay phenotype of cells lacking the 3'-5' exonuclease Rrp6p. Our reporter is capable of performing high-throughput, rapid, and large-scale screens to identify and characterize genetic and chemical perturbations of the major eukaryotic gene expression processes.
authors.forename: Matthew R
authors.initials: MR
authors.lastname: Sorenson
authors.forename: Scott W
authors.initials: SW
authors.lastname: Stevens
mesh_terms: D042002:Chromatin Assembly and Disassembly / Q000235:genetics; D005434:Flow Cytometry; D015966:Gene Expression Regulation, Fungal; D059365:Nonsense Mediated mRNA Decay / Q000235:genetics*; D012322:RNA Precursors; D012326:RNA Splicing / Q000235:genetics*; D020871:RNA Stability / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics
chemical_list: D012322:RNA Precursors
keywords: drug screening; flow cytometry; gene expression; mRNA processing; pre-mRNA splicing