A novel plasmid-based microarray screen identifies suppressors of rrp6Delta in Saccharomyces cerevisiae.
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.
Abruzzi
Katharine
K
Howard Hughes Medical Institute and Department of Biology, Brandeis University, Waltham, MA 02454, USA.
Denome
Sylvia
S
Olsen
Jens Raabjerg
JR
Assenholt
Jannie
J
Haaning
Line Lindegaard
LL
Jensen
Torben Heick
TH
Rosbash
Michael
M
D015536:Down-Regulation; D005095:Exoribonucleases / Q000235:genetics* / Q000378:metabolism*; D063326:Exosome Multienzyme Ribonuclease Complex; D017353:Gene Deletion; D015966:Gene Expression Regulation, Fungal / Q000235:genetics*; D005800:Genes, Fungal / Q000235:genetics; D016153:Genes, Suppressor; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D010957:Plasmids / Q000235:genetics*; D026723:Polyadenylation; D011485:Protein Binding; D020871:RNA Stability; D012331:RNA, Fungal / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012335:RNA, Ribosomal / Q000235:genetics / Q000378:metabolism; D020537:RNA, Small Nucleolar / Q000235:genetics / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000235:genetics* / Q000254:growth & development; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics* / Q000378:metabolism*; D013489:Suppression, Genetic; D013696:Temperature
D012331:RNA, Fungal; D012333:RNA, Messenger; D012335:RNA, Ribosomal; D020537:RNA, Small Nucleolar; D016601:RNA-Binding Proteins; D029701:Saccharomyces cerevisiae Proteins; D005095:Exoribonucleases; D063326:Exosome Multienzyme Ribonuclease Complex; C112610:RRP6 protein, S cerevisiae