Project description:This experiment was designed to investigate the epigenetic basis of sex chromosome dosage compensation. Such mechanisms often operate through epigenetic modifications, including histone modifications, which alter chromatin structure and gene expression. In Drosophila melanogaster, dosage compensation is achieved by upregulating the single X chromosome in males through acetylation of histone H4 at lysine 16 (H4K16ac). We therefore used D. melanogaster as a positive control to validate our approach. In contrast, the mechanism of dosage compensation in Bacillus grandii remains unknown. By profiling H4K16ac in this species, we aim to test whether this histone modification may have been convergently recruited to mediate dosage compensation.
Project description:RNA was extracted from adult male and adult female Drosophila melanogaster with reversed sex-chromosome parent-of-origin (e.g. maternal-X/paternal-Y vs. paternal-X/maternal-Y)
Project description:Drosophila X chromosomes are subject to dosage compensation in males and are known to have a specialized chromatin structure in the male soma. We are interested in how specific chromatin structure change contributes to X chromosome hyperactivity and dosage compensation. We have conducted a global analysis of localize two dosage compensation complex dependent histone marks H4AcK16 and H3PS10 and one dosage compensation complex independent histone mark H3diMeK4 in the genome, especially on X chromosome by ChIP-chip approach in both male and female adult flies. We also probed general genomewide chromatin structure by deep DNA sequencing of sheared ChIP input DNA from male and female adult flies. Chromatin immunoprecipitations were performed in 5-7 day aged adult male and female flies with three histone modification antibodies. ChIP enriched DNA and input DNA was labeled by Cy3 or Cy5 dye separately and hybridized simultaneously to the Drosophila FlyGEM arrays. At least two biological replicates were performed for each antibody and sex. DNA-seq (NIDDK-Drosophila-Illumina-DNASeq) were performed on ChIP-input sheared DNA to check the general chromatin structure of different chromosome.
Project description:Proteomic Analysis (MS/MS) of Drosophila melanogaster mtx2 (Ortholog of CG8004) Heterozygous versus Homozygous Mutants at 2 Days Post-Pupa Formation