A genetic screen for yeast genes induced by sustained osmotic stress.
The budding yeast, Saccharomyces cerevisiae, responds to changes in external osmolarity through the activation of an osmosensing signal transduction pathway. Using lacZ-reporter gene fusions, clonal cell lines were screened for levels of beta-galactosidase activity in the presence or absence of osmotic stress. A screen of 9,000 transformants displayed 663 (7%) gene fusions that were active in rich medium. Each of the transformants were also assayed for gene activity 24 h following a transfer to high osmolarity medium (0.6 M NaCl) and of the 9,000 clonal cell lines, 86 (1%) displayed a decrease in expression, and seven (0.1%) displayed a reproducible increase in gene expression during primary screening. The chromosomal loci of the lacZ insertions were determined, and the gene(s) associated with that site was examined for osmotically induced expression using RNA blot analysis. Five stress-activated genes were analysed by RNA blot: YDL222C, NMD2, PTC7, FAA4 and YRF1. The genes identified by this screen encompass cellular adaptations to stress including signal transduction, protein myristoylation and fatty acid/sphingolipid content in the cell membrane.
Runner
Vanessa M
VM
Natural Science Division, Pepperdine University, 24255 Pacific Coast Highway, Malibu, CA 90263, USA.
Brewster
Jay L
JL
D000214:Acyl Coenzyme A / Q000235:genetics / Q000502:physiology; D048868:Adaptor Proteins, Signal Transducing; D015152:Blotting, Northern; D004265:DNA Helicases / Q000235:genetics / Q000502:physiology; D004268:DNA-Binding Proteins / Q000235:genetics / Q000502:physiology; D015966:Gene Expression Regulation, Fungal / Q000502:physiology*; D005800:Genes, Fungal / Q000502:physiology*; D016681:Genome, Fungal; D047908:Intracellular Signaling Peptides and Proteins; D016254:Mutagenesis, Insertional; D009994:Osmolar Concentration; D010749:Phosphoprotein Phosphatases / Q000235:genetics / Q000502:physiology; D016133:Polymerase Chain Reaction; D011494:Protein Kinases; D054648:Protein Phosphatase 2; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000502:physiology; D015398:Signal Transduction / Q000235:genetics / Q000502:physiology; D015534:Trans-Activators / Q000235:genetics / Q000502:physiology; D014170:Transformation, Genetic
D000214:Acyl Coenzyme A; D048868:Adaptor Proteins, Signal Transducing; D004268:DNA-Binding Proteins; D047908:Intracellular Signaling Peptides and Proteins; C092470:NMD2 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D015534:Trans-Activators; D011494:Protein Kinases; C102035:YPD1 protein, S cerevisiae; C433364:PTC7 protein, S cerevisiae; D010749:Phosphoprotein Phosphatases; D054648:Protein Phosphatase 2; D004265:DNA Helicases; C116427:Y'-helicase protein 1, S cerevisiae; C090206:FAA4 protein, S cerevisiae