Chemical-genetic profile analysis in yeast suggests that a previously uncharacterized open reading frame, YBR261C, affects protein synthesis.
BACKGROUND
Functional genomics has received considerable attention in the post-genomic era, as it aims to identify function(s) for different genes. One way to study gene function is to investigate the alterations in the responses of deletion mutants to different stimuli. Here we investigate the genetic profile of yeast non-essential gene deletion array (yGDA, approximately 4700 strains) for increased sensitivity to paromomycin, which targets the process of protein synthesis.


RESULTS
As expected, our analysis indicated that the majority of deletion strains (134) with increased sensitivity to paromomycin, are involved in protein biosynthesis. The remaining strains can be divided into smaller functional categories: metabolism (45), cellular component biogenesis and organization (28), DNA maintenance (21), transport (20), others (38) and unknown (39). These may represent minor cellular target sites (side-effects) for paromomycin. They may also represent novel links to protein synthesis. One of these strains carries a deletion for a previously uncharacterized ORF, YBR261C, that we term TAE1 for Translation Associated Element 1. Our focused follow-up experiments indicated that deletion of TAE1 alters the ribosomal profile of the mutant cells. Also, gene deletion strain for TAE1 has defects in both translation efficiency and fidelity. Miniaturized synthetic genetic array analysis further indicates that TAE1 genetically interacts with 16 ribosomal protein genes. Phenotypic suppression analysis using TAE1 overexpression also links TAE1 to protein synthesis.


CONCLUSIONS
We show that a previously uncharacterized ORF, YBR261C, affects the process of protein synthesis and reaffirm that large-scale genetic profile analysis can be a useful tool to study novel gene function(s).
Alamgir
Md
M
Department of Biology, Carleton University, 1125 Colonel By Drive, Ottawa, K1S 5B6, Canada. malamgir@connect.carleton.ca
Eroukova
Veronika
V
Jessulat
Matthew
M
Xu
Jianhua
J
Golshani
Ashkan
A
D020869:Gene Expression Profiling / Q000379:methods*; D008780:Methyltransferases / Q000235:genetics* / Q000502:physiology; D016366:Open Reading Frames; D010303:Paromomycin / Q000494:pharmacology; D014176:Protein Biosynthesis / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000502:physiology; D017384:Sequence Deletion; D049490:Systems Biology
D029701:Saccharomyces cerevisiae Proteins; D010303:Paromomycin; D008780:Methyltransferases; C535246:TAE1 protein, S cerevisiae