Project description:We have used microarrays to identify DSX occupancy signal in adult female fatbody using the DamID protocol. We have performed DamID-chip on adult female fatbody with three biological replicates.
Project description:We have adapted the DamID protocol for use with high throughput sequencing. We have used DamID to identify the positions within the Drosophila genome where the transcription factor DSX is bound. We sequenced DpnI-digested genomic DNA from fly tissues containing UAS-Dam (control) or UAS-Dam-DsxF or UAS-Dam-DsxM. We have performed DamID-seq on adult male and female fatbody and on ovary. We used two biological replicates for each tissue and sex.
Project description:Drosophila melanogaster undergoes a complete metamorphosis, during which time the larval male and female forms transition into sexually dimorphic, reproductive adult forms. To understand this complex morphogenetic process at a molecular-genetic level, we performed whole genome microarray analyses. In somatic tissues at 48 hour After Puparium Formation (APF), 173 sex-biased transcripts that likely function downstream of the doublesex (dsx) branch of the sex determination hierarchy were identified. The mode of regulation of the sex-specific isoforms of DSX (DSX-F and DSX-M) was examined. It was determined that for most downstream targets, DSX-F and DSX-M regulate gene expression in the same manner, but that one isoform acts as a more potent regulator. Keywords: wild type; genetic modification All microarrays were dual channel with direct comparisons of male versus female or wild type versus mutant. All samples consist of whole body pupae collected at 48 hour After Puparium Formation (APF). For each experiment, four biological replicates were analyzed in a dye-swap design.
Project description:TCDD is an environmental contaminant that elicits a number of hepatic effects including fat accumulation, inflammation, and fibrosis that can progress to hepatocellular carcinoma. RNA-Seq and targeted metabolomics were integrated with complementary dioxin response element (DRE) location and aryl hydrocarbon receptor (AhR) ChIP-Seq data to further investigate the hepatotoxicity of TCDD. Our integrative analysis identified changes similar to the Warburg effect observed in cancer cells, including pyruvate kinase isoform switching (PKM1 to PKM2), and an increase in the glutaminase (GLS1) GAC:KGA isoform ratio. Consequently, metabolites are redirected towards the pentose phosphate pathway, serine biosynthesis, and glutaminolysis. We propose that the effects of TCDD on central carbon and amino acid metabolism represents AhR-mediated hepatic metabolic reprogramming in order to increase NADPH production as an oxidative stress counter-measure.
Project description:Drosophila melanogaster undergoes a complete metamorphosis, during which time the larval male and female forms transition into sexually dimorphic, reproductive adult forms. To understand this complex morphogenetic process at a molecular-genetic level, we performed whole genome microarray analyses. In somatic tissues at 48 hour After Puparium Formation (APF), 173 sex-biased transcripts that likely function downstream of the doublesex (dsx) branch of the sex determination hierarchy were identified. The mode of regulation of the sex-specific isoforms of DSX (DSX-F and DSX-M) was examined. It was determined that for most downstream targets, DSX-F and DSX-M regulate gene expression in the same manner, but that one isoform acts as a more potent regulator. Keywords: wild type; genetic modification
Project description:We have adapted the DamID protocol for use with high throughput sequencing. We have used DamID to identify the positions within the Drosophila genome where the transcription factor DSX is bound. We sequenced DpnI-digested genomic DNA from fly tissues containing UAS-Dam (control) or UAS-Dam-DsxF or UAS-Dam-DsxM.