Project description:This study newly identified Tripelennamine (TA) as an inhibitor of yeast meiosis and sporulation. To examine if and how exposure of sporulating yeast cells to TA influences the viability of the cells a homozygous deletion pool experiment was conducted.
Project description:A pool of 3633 tagged heterozygous transposon disruption mutants underwent haploinsufficiency profiling in different media conditions (YPD, SC, YNB, and SLAD) to identify slow growing strains in these conditions.
Project description:A pool of 3633 tagged heterozygous transposon disruption mutants underwent haploinsufficiency profiling in the presence of different synthetic compounds to identify their cellular targets
Project description:RSC is a growth essential ATP-dependent chromatin-remodeling complex of Saccharomyces cerevisiae. Nps1/Sth1 is the ATPase subunit of the complex. A temperature-sensitive mutant allele of NPS1, nps1-13 and the null mutation of the RSC2 or RSC7 gene showed growth defect on a medium containing non fermentable carbon source, such as lactate or ethanol-glycerol (YPEG), suggested a possibility that RSC plays a role on mitochondria function. We used microarrays to compare the global gene expression profiles between the wild type and nps1-13 mutant under respiratory condition. WT (BY4743) and nps1-13 strains pre-grown in YPD medium were inoculated in YPEG medium for RNA extraction and hybridization on Affymetrix microarrays. In order to obtain more global insights into the role of RSC under respiratory conditions, we perfomed the genome-wide expression analysis of nps1-13 in YPEG medium.
Project description:Histone acetylation and deacetylation are among the principal mechanisms by which chromatin is regulated during transcription, DNA silencing, and DNA repair. We analyzed patterns of genetic interactions uncovered during comprehensive genome-wide analyses in yeast to probe how histone acetyltransferase (HAT) and histone deacetylase (HDAC) protein complexes interact. The genetic interaction data unveil an underappreciated role of HDACs in maintaining cellular viability, and led us to show that deacetylation of the histone variant Htz1p at lysine 14 is mediated by Hda1p. Studies of the essential nucleosome acetyltransferase of H4 (NuA4) revealed acetylation-dependent protein stabilization of Yng2p, a potential nonhistone substrate of NuA4 and Rpd3C, and led to a new functional organization model for this critical complex. We also found that DNA double-stranded breaks (DSBs) result in local recruitment of the NuA4 complex, followed by an elaborate NuA4 remodeling process concomitant with Rpd3p recruitment and histone deacetylation. These new characterizations of the HDA and NuA4 complexes demonstrate how systematic analyses of genetic interactions may help illuminate the mechanisms of intricate cellular processes. Keywords: genetic modification The 44 datasets in this Series profiled the genome-wide genetic interactions for query genes encoding either HAT and HDAC catalytic subunits or subunits of the associated protein complexes. Of the 32 query genes, 5 were essential and were tested as temperature-sensitive (ts) alleles at three or more temperatures. (ESA1 was also tested as a hypomorphic allele.) The other query genes were tested as null deletion alleles derived from the Yeast Knockout strain collection.
Project description:Non-coding RNAs (ncRNAs), including the more recently identified Stable Unannotated Transcripts (SUTs) and Cryptic Unstable Transcripts (CUTs), are increasingly being shown to play pivotal roles in the transcriptional and post-transcriptional regulation of genes in eukaryotes. Here, we carried out a large-scale screening of ncRNAs in Saccharomyces cerevisiae, and provide evidence for SUT and CUT function. Phenotypic data on 372 ncRNA deletion strains in 23 different growth conditions were collected, identifying ncRNAs responsible for significant cellular fitness changes. Transcriptome profiles were assembled for 18 haploid ncRNA deletion mutants and 2 essential ncRNA heterozygous deletants. Guided by the resulting RNA-seq data we analysed the genome-wide dysregulation of protein coding genes and non-coding transcripts.
Project description:The complete pool of barcoded essential heterozygous diploid deletion strains of S. cerevisiae were screened with 20 compounds from the Chembridge NOVACore chemical library to identify gene deletions that confer sensitivity to each compound.
Project description:The complete pool of barcoded homozygous and essential heterozygous diploid deletion strains of S. cerevisiae were screened with 3-nitroso-imidazo[1,2-a]pyridines and -pyrimidines to identify gene deletions that confer sensitivity to each compound.
Project description:Reactive oxygen species, generated in vivo or exogenously encountered, constantly challenge living organisms. Oxidation of polyunsaturated fatty acids (PUFA), which are susceptible to oxidant attack, can lead to initiation of lipid peroxidation and in turn rapid production of toxic lipid hydroperoxides. Eukaryotic microorganisms such as Saccharomyces cerevisiae can survive harsh industrial conditions that contain high levels of the PUFA linoleic acid and its oxidised derivative, linoleic acid hydroperoxide (LoaOOH). The precise signalling and response mechanisms induced by yeast to overcome lipid hydroperoxide stress are ill understood. We used genome-wide microarrays to investigate the changes in gene expression of S. cerevisiae to LoaOOH-induced oxidative stress. S. cerevisiae (BY4743) were exposed to an arresting concentration of LoaOOH (75 M-BM-5M) for 1 hr to induce oxidative stress. Yeast treated with an equivalent volume of solvent (methanol) were used as a control. Following treatment conditions, total RNA was extracted from LoaOOH-treated or control yeast and hybridised onto Affymetrix microarrays.
Project description:This study used untargeted proteomics to compare blood proteomic profiles in two groups of adults that differed widely in lifestyle habits. The goal was to identify a core list of proteins that were either upregulated or downregulated based on adherence to recommended lifestyle habits in adults. A total of 52 subjects in the lifestyle group (LIFE) (28 males, 24 females) and 52 in the control group (CON) (27 males, 25 females) participated in this cross-sectional study. Age, education level, marital status, and height did not differ significantly between LIFE and CON groups. The LIFE and CON groups differed markedly in body composition and fat mass-related anthropometric measurements including waist circumference, body mass index (BMI), sagittal abdominal diameter (SAD), and body composition (body fat percentage and fat mass index or FMI) (p<0.001). LIFE and CON groups were also widely disparate in physical activity patterns and maximal aerobic fitness, dietary intake patterns, disease risk factor prevalence, blood measures of inflammation, triglycerides, HDL-cholesterol, glucose, and insulin, weight-adjusted leg/back and handgrip strength, and mood states. The proteomics analysis showed strong group differences for 39 of 725 proteins identified in the dried blood spot samples. Of these, 18 were downregulated in the LIFE group and collectively indicated a lower innate immune activation signature. A total of 21 proteins were upregulated in the LIFE group and supported greater lipoprotein metabolism and HDL remodeling. Lifestyle-related habits and biomarkers were probed and the variance (>50%) in downregulated and upregulated proteins was best explained by group contrasts in indicators of body composition and visceral fat including FMI and SAD. This cross-sectional study established that a relatively small number of upregulated and downregulated proteins are associated with good lifestyle habits. A targeted “lifestyle” proteomic panel based on these data could be used in future studies to determine the efficacy of various prevention and treatment strategies.