Disruption and phenotypic analysis of six open reading frames from the left arm ofSaccharomyces cerevisiae chromosome VII

Abstract

Six open reading frames (ORFs) from Saccharomyces cerevisiae chromosome VII were deleted using the kanMX4 module and the long‐flanking homology‐PCR replacement strategy in at least two different backgrounds. Among these ORFs, two of them (YGL100w and YGL094c) are now known genes which encode well‐characterized proteins (Seh1p, a nuclear pore protein, and Pan2p, a component of Pab1p‐stimulated poly(A) ribonuclease, respectively). The other four ORFs (YGL101w, YGL099w, YGL098w and YGL096w) code for proteins of unknown function, although the protein encoded by YGL101w has a strong similarity to the hypothetical protein Ybr242p. Gene disruptions were performed in diploid cells using the KanMX4 cassette, and the geneticin (G418)‐resistant transformants were checked by PCR. Tetrad analysis of heterozygous deletant strains revealed that YGL098w is an essential gene for vegetative growth in three backgrounds, whereas the other five genes are non‐essential, although we have found some phenotypes in one of them. YGL099wΔ strain did not grow at all at 15°C and showed a highly impaired sporulation and a significantly lower mating efficiency. The other three deletants did not reveal any significant differences with respect to their parental strains in our basic phenotypic tests. Copyright © 2000 John Wiley & Sons, Ltd.

Mutants defective in secretory/vacuolar pathways in the EUROFAN collection of yeast disruptants

“…It encodes a protein carrying a potential ATP/GTP binding motif that has numerous homologues in databases. The null mutant displayed slow growth and abnormal morphology (Lillo et al, 2000). Our data suggest that modifications in the wall of these cells may arise from impaired glycosylation.…”

Towards the prediction of essential genes by integration of network topology, cellular localization and biological process information

“…Analyzing these genes, we found that 9 genes have been previously demonstrated to be essential ( YHR165C , YHR089C , YHR052W , YCR042C , YDR320C-A , YHR169W , YKL138C-A , YGL106W and YHR099W ) and other 14 genes ( YGR252W , YHR027C , YOL012C , YNL147W , YGL100W , YNL096C , YOL148C , YFL007W , YOL145C , YBR111W-A , YNL055C , YHR216W , YBL071W-A and YHR039C-A ) have been previously demonstrated to be non-essential by other investigators through small-scale gene deletion experiments in functional characterization studies [25-36] (Table 1). Among non-essential genes, 10 genes ( YGR252W , YHR027C , YOL012C , YNL147W , YNL096C , YOL148C , YOL145C , YBR111W-A , YNL055C and YHR039C-A ) have been shown to impair substantially the growth of S. cerevisiae when they are completely deleted [33,36-40], whereas the 4 remaining non-essential genes ( YGL100W , YFL007W , YHR216W and YBL071W-A ) have been shown not to affect the growth phenotype of yeast when they are deleted [34,35,41,42]. Although roughly 1/3 of the these genes predicted to be essential have been previously classified as non-essential, the complete deletions of most of them have been shown to severely reduce the fitness of organisms [33,36-40], suggesting that our predictor, even when directly contradicted by these experimental findings, can nonetheless identify genes important to cellular function.…”