Project description:Genome wide map of heterochromatin state in fission yeast Schizosaccharomyces pombe via 4 different strains Examination of a single histone modification in 4 different fission yeast strains
Project description:We used massively parallel sequencing to discover and characterize small RNAs (sRNAs) from fission yeast Schizosaccharomyces japonicus. We found that, unlike in related S. pombe, a substantial fraction of sRNAs maps to transposons, both telomeric and centromeric.
Project description:The phosphorylation of proteins modulates various functions of proteins and plays an important role in regulation of cell signaling. In the recent years, the label-free quantitative (LFQ) phos-phoproteomics has become the powerful tool to analyze the phosphorylation of proteins within the complex samples. Despite the great progress, the studies of protein phosphorylations are still limited in throughput, robustness, and reproducibility, hampering analyses that involve multiple perturbations, such as those needed to follow the dynamics of phosphoproteomes. To address these challenges, we introduce here the LFQ phosphoproteomics workflow that is based on Fe-IMAC phosphopeptide enrichment followed by strong anion exchange (SAX) and porous graphitic carbon (PGC) fractionation strategies. We applied this workflow to analyze the whole-cell phosphoproteome of the fission yeast Schizosaccharomyces pombe. Using the strategy, we identified 8353 phosphosites from which 1274 were newly identified. This provides the sig-nificant addition to the S. pombe phosphoproteome. Results of our study highlight that combining of PGC and SAX fractionation strategies substantially increases the robustness and specificity of LFQ phosphoproteomics. Overall, the presented LFQ phosphoproteomics workflow opens the door for studies that would get better insight into the complexity of the protein kinase functions of the fission yeast S. pombe.
Project description:We used massively parallel sequencing to discover and characterize small RNAs (sRNAs) from fission yeast Schizosaccharomyces japonicus. We found that, unlike in related S. pombe, a substantial fraction of sRNAs maps to transposons, both telomeric and centromeric. small RNA library from total RNA isolations from Schizosaccharomyces japonicus
Project description:Nitric oxide being a versatile molecule inside biological systems, from being both a cell signaling molecule to a potent stress agent, has significant effect in the transcriptional response in fission yeast. We have used fission yeast microarrays to identify cellular targets of Nitric Oxide (NO) and to further understand the cellular mechanism of NO action. We report the change in the global gene expression profile response to NO in S. pombe cells
Project description:RNA interference (RNAi) is a gene silencing mechanism conserved from fungi to mammals. Small interfering RNAs are products and mediators of the RNAi pathway and act as specificity factors in recruiting effector complexes. The Schizosaccharomyces pombe genome encodes one of each of the core RNAi proteins, Dicer, Argonaute and RNA-dependent RNA polymerase (dcr1, ago1, rdp1). Even though the function of RNAi in heterochromatin assembly in S. pombe is established, its role in controlling gene expression is elusive. Here, we report the identification of small RNAs mapped anti-sense to protein coding genes in fission yeast. We demonstrate that these genes are up-regulated at the protein level in RNAi mutants, while their mRNA levels are not significantly changed. We show that the repression by RNAi is not a result of heterochromatin formation. Thus, we conclude that RNAi is involved in post-transcriptional gene silencing in S. pombe.