A novel plasmid-based microarray screen identifies suppressors of rrp6Delta in Saccharomyces cerevisiae

Mol Cell Biol. 2007 Feb;27(3):1044-55. doi: 10.1128/MCB.01299-06. Epub 2006 Nov 13.

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 rrp6Delta 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 rrp6Delta strains at 37 degrees 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 rrp6Delta strains. Microarray analyses of gene expression in rrp6Delta strains and a number of suppressor strains support this hypothesis.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Down-Regulation
  • Exoribonucleases / genetics*
  • Exoribonucleases / metabolism*
  • Exosome Multienzyme Ribonuclease Complex
  • Gene Deletion*
  • Gene Expression Regulation, Fungal / genetics*
  • Genes, Fungal / genetics
  • Genes, Suppressor*
  • Oligonucleotide Array Sequence Analysis / methods*
  • Plasmids / genetics*
  • Polyadenylation
  • Protein Binding
  • RNA Stability
  • RNA, Fungal / genetics
  • RNA, Fungal / metabolism
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • RNA, Ribosomal / genetics
  • RNA, Ribosomal / metabolism
  • RNA, Small Nucleolar / genetics
  • RNA, Small Nucleolar / metabolism
  • RNA-Binding Proteins / metabolism
  • Saccharomyces cerevisiae / cytology
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae / growth & development
  • Saccharomyces cerevisiae Proteins / genetics*
  • Saccharomyces cerevisiae Proteins / metabolism*
  • Suppression, Genetic
  • Temperature

Substances

  • RNA, Fungal
  • RNA, Messenger
  • RNA, Ribosomal
  • RNA, Small Nucleolar
  • RNA-Binding Proteins
  • Saccharomyces cerevisiae Proteins
  • Exoribonucleases
  • Exosome Multienzyme Ribonuclease Complex
  • RRP6 protein, S cerevisiae