Affymetrix custom RNA microarray for WT and jhd2-delete strains in terminal spores (20h)
Ontology highlight
ABSTRACT: RNA transcript signals were profiled in WT (MMY718) and jhd2M-bM-^HM-^F (MMY1879) terminally sporulated cultures (20h of sporulation) using Affymetrix high resolution tiling microarrays. Total RNA was isolated, biotin labelled, and hybridized to custom Affymetrix Scerevisiae_tlg tiling arrays. The array contains 25nt probes covering the Watson strand of the yeast genome with 8nt tiling resolution, and another set of 25nt probes covering the Crick strand with 8nt resolution, with a 4nt offset from the Watson probes. This gives an overall tiling resolution of 4bp with a total of 6.5 million probes, and also allows detection of strand-specific RNA signals.
Project description:Transfer RNA (tRNA) modifications enhance the efficiency, specificity and fidelity of translation in all organisms. The anticodon modification mcm5s2U34 is required for normal growth and stress resistance in yeast; mutants lacking this modification have numerous phenotypes. Mutations in the homologous human genes are linked to neurological disease. The yeast phenotypes can be ameliorated by overexpression of specific tRNAs, suggesting that the modifications are necessary for efficient translation of specific codons. We determined the in vivo ribosome distributions at single codon resolution in yeast strains lacking mcm5s2U. We found accumulations at AAA, CAA, and GAA codons, suggesting that translation is slow when these codons are in the ribosomal A site, but these changes appeared too small to affect protein output. Instead, we observed activation of the GCN4-mediated stress response by a non- canonical pathway. Thus, loss of mcm5s2U causes global effects on gene expression due to perturbation of cellular signaling. WT yeast and mutants lacking anticodon tRNA modifications were grown in YPD, and subjected to ribosome footprint profiling (ribo-seq) and RNA-seq of poly-A selected RNA. Dataset contains biological replicates for WT, Δncs6 and Δuba4. Technical replicates were also performed for all RNA-seq datasets (using a different poly-A selection method).
Project description:Using chromatin immunoprecipitation and high-resolution tiling arrays covering the human genome, we render a genome-wide map of p73 DNA binding sites in ME180 human cervical carcinoma cells.
Project description:To identify meiotic non-coding RNAs, a DNA strand-specific transcript profiling experiment using cultures of Saccharomyces cerevisiae in YPD, YPA and SPII medium and tiling microarrays (GeneChips) covering both strands of the strain S288c complete genome was carried out.
Project description:Human DNA replication relies on the activation of thousands of origins distributed along the genome. Their organization and the functional significance of their distribution remains unknown. To map large number of origins in human cells, we have localized the distribution of short nascent DNA using a high-resolution DNA tiling array platform covering 33.9Mb. Results with three different cell lines reveal that origins are very closely spaced (average interval 3-5kb), and that their positions are largely conserved among the cell lines studied. Origins are non-randomly distributed, being preferentially enriched at the 5’-end of expressed genes and at evolutionary intergenic sequences. In MCF7 cells, a strong correlation is also found between origin positioning and histone H3K4me3 and of PolII binding to chromatin. This study provides a large scale view of the organization of DNA replication origins in human chromosomes and suggests a link between this distribution and underlying chromatin features related to chromosome structure, and gene expression Keywords: nascent strand DNA, histone, Pol-II ChIP-chip. To map initiation sites for DNA replication and the position of both H3K4Me3 and Pol-II chromatin binding sites in human cell lines on a custom made high density tiling DNA microarray covering 33.9 Mb of the human genome.
Project description:High-resolution transcriptional profiling of E. coli across nine timepoints of growth in rich media (LB). Samples collected from lag phase to stationary phase of growth. High-resolution tiling array to detect conditional operon isoforms. Custom Agilent array designed to detect condition-specific transcriptional isoforms. Array designed for E coli K12 MG1655 genome. Tiled using 23bp sliding window. Includes >10,000 probes surrounding predicted operon break sites at 6 bp resolution.
Project description:The human MHC is a paradigm for genomics, showing striking association with disease but functional variants remain largely unresolved. Using an original hybrid microarray (containing tiling and junction probes) for the MHC and accounting for known sequence diversity, we have drawn the first high-resolution, strand-specific transcriptional map of the MHC, defining differences in gene expression for three common haplotypes associated with autoimmune disease. In total, 6% of the MHC is transcribed with one transcript per 1.4kb, including previously unrecognized intergenic transcription. The distributions of differentially expressed probes and polymorphisms between haplotypes are significantly correlated, arguing for cis effects. Haplotype-specific transcription involved 96 differentially expressed genes, including ZFP57, which was validated in a cohort of healthy volunteers, while 526 exons show haplotypic differences. We also find splicing events are significantly more extensive in the MHC than in the rest of the genome. This study marks a new step in immunogenetics. The results files (.bedgraph and .gff) contain tiling data for both the shared-path and the alternate paths (shared and haplotype-specific) , defining transcriptional activity across the entire MHC region in each sample.
Project description:Deep sequencing was implemented to study the transcriptional landscape of Mycobacterium avium TMC724. High-resolution transcriptome analysis identified the transcription start points for 652 genes. One third of these coincided with the start codons and therefore belong to leaderless transcripts, whereas the rest of the transcripts had 5' UTRs with the mean length of 83 nt. In addition, the 5' UTRs of 6 genes contained SAM-IV and Ykok types of riboswitches. 87 antisense RNAs and 9 intergenic small RNAs were mapped. Four of the revealed intergenic small RNAs, including igMAV_1034-1035 expressed at a very high level, have no homologs in M. tuberculosis, whilst M. avium lacks several intergenic sRNAs present in M. tuberculosis. Among those, MTS479 and MTS1338 are of special interest due to their possible implication in pathogenesis. Elucidation of differences in the repertoire of intergenic sRNAs between the two mycobacterial species may improve our understanding of mycobacterial diseases pathogenesis. Transcriptional profile of Mycobacterium avium TMC724, grown at 37M-BM-0C in Dubos broth until mid-logarithmic growth phase
Project description:HighRes-CGH arrays that utilize ≈385,000 distinct oligonucleotide probes to cover chromosome 22 at 85 bp resolution (tiling path step size) were designed and synthesized as previously described (Urban et al., Proc Natl Acad Sci U S A. 2006;103(12):4534-9; see also GSE4240 record). Here, two healthy individuals were studied using the same experimental protocols as in Urban et al. Keywords: high-resolution comparative genome hybridization using oligonucleotides
Project description:Genome-scale DNA methylation profiles at nucleotide resolution covering the vast majority of CpG islands and a representative sampling of conserved non-coding elements, transposons and other genomic features generated using high-throughput reduced representation bisulfite sequencing (RRBS) for murine hematopoietic stem cells during ontongeny and after enforced prolferative history. Analysis of hematopoietic stem cell methylation during ontogeny and after undergoing enforced proliferative history
Project description:The sea urchin S. purpuratus is a model organism for study of the genomic control circuitry underlying embryonic development. We examined the complete repertoire of genes expressed in the S. purpuratus embryo, up to late gastrula stage, by means of high-resolution custom tiling arrays covering the whole genome. We detected complete spliced structures even for genes known to be expressed at low leveles in only a few cells. At least 11,000 to 12,000 genes are used in embryogenesis. These include most of the genes encoding transcription factors and signaling proteins, as well as some classes of general cytoskeletal and metabolic proteins, but only a minor fraction of genes encoding immune functions and sensory receptors. Thousands of small asymmetric transcripts of unknown function were also detected in intergenic regions throughout the genome. The tiling array data were used to correct and authenticate several thousand gene models during the genome annotation process. Keywords: high-resolution tiling array, sea urchin, embryo Keywords: other