A Novel Plasmid-Based Microarray Screen Identifies Suppressors of rrp6 Δ in Saccharomyces cerevisiae

Abstract

ABSTRACT Genetic screens in Saccharomyces cerevisiae provide novel information about interacting genes and pathways. We screened for high-copy-number suppressors of a strain with the gene encoding the nuclear exosome component Rrp6p deleted, with either a traditional plate screen for suppressors of rrp6 Δ temperature sensitivity or a novel microarray enhancer/suppressor screening (MES) strategy. MES combines DNA microarray technology with high-copy-number plasmid expression in liquid media. The plate screen and MES identified overlapping, but also different, suppressor genes. Only MES identified the novel mRNP protein Nab6p and the tRNA transporter Los1p, which could not have been identified in a traditional plate screen; both genes are toxic when overexpressed in rrp6 Δ strains at 37°C. Nab6p binds poly(A) + RNA, and the functions of Nab6p and Los1p suggest that mRNA metabolism and/or protein synthesis are growth rate limiting in rrp6 Δ strains. Microarray analyses of gene expression in rrp6 Δ strains and a number of suppressor strains support this hypothesis.