Project description:Sequencing of mononucleosomal DNA during asynchronous mitosis and 0, 3 and 5 hours into meiosis in Schizosaccharomyces pombe. Two samples from mononucleosomal DNA from asynchronous mitosis (haploid 972 h- and diploid pat1.114) and three samples from 0, 3 and 5 hours into meiosis (from diploid pat1.114) were sequenced (Illumina Genome Analyzer IIx) using the single-end read protocol.
Project description:Opi10 is the S. pombe homolog of human Hikeshi, which imports Hsp70s into the nucei during the heat shock. We compared the gene expression of the S. pombe opi10+ and opi10- strains before, during and after the heat shock. RNA were extracted from the S. pombe opi10+ and opi10- strains cultured at 30C, heat treated for 1 h at 43C, and then cultred for 1 and 3 h at 30C, and analyzed using Affymerix DNA microarrays.
Project description:Schizosaccharomyces pombe Rad3 checkpoint kinase and its human ortholog ATR are essential for maintaining genome integrity in cells treated with genotoxins that damage DNA or arrest replication forks. Rad3 and ATR also function during unperturbed growth, although the events triggering their activation and their critical functions are largely unknown. Here, we use ChIP-on-chip analysis to map genomic loci decorated by phosphorylated histone H2A (gH2A), a Rad3 substrate that establishes a chromatin-based recruitment platform for DNA repair/checkpoint proteins. Our data showed that gH2A marks a diverse array of genomic features during S-phase, including natural replication fork barriers and a fork breakage site, retrotransposons, heterochromatin in the centromeres and telomeres, and ribosomal RNA (rDNA) repeats. The enrichment of gH2A at these sites was confirmed by multiple ChiP-qPCR experiments. Overall design: Phosphorylated Histone H2A was ChIPed with anti-gammaH2A antibody vs. Input(whole cell extract) in S. pombe cells synchronized in S-phase.
Project description:A minimal cell cycle control module is driving the cell cycle in these S. Pombe cells. During the experiment the cells are arrested or released to cycle through the different phases of the cell cycle. Samples were collected at the indicated times points from arrested and cycling cells and expression of genes were measured by microarrays on an Affymetrix platform.
Project description:Hrp3_Purification from Schizosaccharomyces pombe 972h- Eukaryotic genome is composed of repeating units of nucleosomes to form chromatin arrays. A canonical gene is marked by nucleosome free region (NFR) at its 5’ end followed by uniformly spaced arrays of nucleosomes. In fission yeast we show both biochemically and in vivo that both Hrp1 and Hrp3 are key determinants of uniform spacing of genic arrays.
Project description:Tristetraprolin is a vertebrate CCCH tandem zinc finger protein that can bind to and destabilize certain mRNAs containing AU-rich element binding sites. zfs1 is the single gene in the fission yeast, Schizosaccharomyces pombe, that encodes a protein containing the critical features of the tristetraprolin zinc finger domain. zfs1 has been linked to pheromone signal transduction control and to the coordination of mitosis, but no biological function has been ascribed to the zfs1 protein. Through a functional genomics approach we compared transcript levels in wild-type and zfs1-deficient S. pombe strains; those elevated in the zfs1-deficient strain were examined for the presence of potential tristetraprolin-like binding sites. One such potential target transcript was encoded by arz1, a gene encoding a protein of unknown function that contains armadillo repeats. arz1 mRNA decay was inhibited in the zfs1-deficient strain when it was expressed under the control of a thiamine-repressible promoter. Mutations within one AU-rich element present in the arz1 3’-untranslated region protected this transcript from zfs1-promoted decay, whereas mutating another potential binding site had no effect. Binding assays confirmed a direct interaction between zfs1 and arz1 mRNA-based probes; this interaction was eliminated when key residues were mutated in either zfs1 zinc finger. zfs1 and its targets in S. pombe represent a useful model system for studies of zinc finger protein/AU-rich element interactions that result in mRNA decay. Keywords: knockout comparison, steady-state analysis Overall design: S. pombe wild-type and zfs1-deficient strains were grown to 1.0 A600 in EMM+5S at 30°C for steady state analysis. Total cellular RNA was prepared from four replicate cultures.