Chromatin immunoprecipitation of histone H3 and several methylated H3 variants in Caenorhabditis elegans
Ontology highlight
ABSTRACT:
ORGANISM(S): Caenorhabditis elegans
SUBMITTER: Thomas Down
PROVIDER: E-TABM-623 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
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2016-03-31 | PXD003583 | Pride
Project description:Histone H3 lysine27-to-methionine (H3K27M) gain-of-function mutations occur in highly aggressive pediatric gliomas. Here, we establish a Drosophila animal model for the pathogenic histone H3K27M mutation and show that its overexpression resembles Polycomb repressive complex 2 (PRC2) loss-of-function phenotypes, causing de-repression of PRC2 target genes and developmental perturbations. Similarly, a H3K9M mutant depletes H3K9 methylation levels and suppresses position-effect variegation in various Drosophila tissues. The histone H3K9 demethylase KDM3B/JHDM2 associates with H3K9M nucleosomes and its overexpression in Drosophila results in loss of H3K9 methylation levels and heterochromatic silencing defects. Here we establish histone lysine-to-methionine mutants as robust in vivo tools for inhibiting methylation pathways that also function as biochemical reagents for capturing site-specific histone-modifying enzymes, thus providing molecular insight into chromatin-signaling pathways. RNA-seq of wing imaginal discs expressing either H3.3WT-FLAG-HA or H3.3K27M-FLAG-HA.
2014-09-02 | E-GEOD-59891 | biostudies-arrayexpress
Project description:We used gene expression profiling to address several specific questions that arose in a study of repair of ultraviolet C radiation in C elegans, as well as to generate hypotheses regarding the possible mechanism(s) of decreased DNA repair observed in old adults in that study. This analysis was performed in order to analyze gene expression in the strain (JK1107) and experimental conditions that we used for our DNA repair studies. The supplementary file GSE4766_Resolver_all_data.txt includes Resolver generated fold-changes and p values based on ratios built in Rosetta Resolver as described in the Rosetta Biosoftware Technical Note (Lee Weng, 2004), Data processing and analysis methods in the Rosetta Resolver system (http://www.rosettabio.com/tech/default.htm). Experiment Overall Design: Gene expression data for glp-1 (strain JK1107) nematodes were obtained by comparing mRNA expression level in replicate pools of 2000-3000 mixed-stage embryos, young (1-day) adults, and old (6-day) adults raised at 25ºC. Three replicate pools of embryo and young adult samples were analyzed, but only two replicate pools of 6-day adults were analyzed due to low RNA recovery.
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