Genome-wide analysis of dFOXO binding sites in Drosophila
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ABSTRACT: To understand how reduced insulin/IGF-1 signaling extends Drosophila lifespan through its downstream transcription factor dFOXO. We conducted ChIP analysis with a dFOXO antibody followed by Illumina high-throughput sequencing from chico heterozygous mutants, which are long-lived and normal sized, and from adult flies with ablated insulin producing cells (IPCs), which are also long-lived. dFOXO bound at promoters of 273 genes common among these genotypes, thus potentially enriching for shared factors in control of aging.
Project description:To understand how reduced insulin/IGF-1 signaling extends Drosophila lifespan through its downstream transcription factor dFOXO. We conducted ChIP analysis with a dFOXO antibody followed by Illumina high-throughput sequencing from chico heterozygous mutants, which are long-lived and normal sized, and from adult flies with ablated insulin producing cells (IPCs), which are also long-lived. dFOXO bound at promoters of 273 genes common among these genotypes, thus potentially enriching for shared factors in control of aging. Two replicates were sequenced from chico heterozygous mutants and IPC ablated flies.
Project description:dFOXO targets in adult Drosophila melanogaster females, and the effect of insulin signalling and stress on binding. The experimets determined the binding locations of dFOXO in the whole adult female fly using ChIP-chip. The protocol was validated using mock conditions: pre-immune serum or IP on chromatin from foxo null flies. The response of this binding to stress induced by treatment of flies with paraquat or by their exposure to starvation, as well as the response to an insulin-signalling-reducing genetic manipulation (over-expression of dominant negative form of the insulin receptor), was determined.
Project description:Transcriptional analysis of dInR and dfoxo epistasis in Drosophila melanogaster. The experiment was performed to examine which parts of the transcriptional response to a reduction in insulin signalling in an adult female fly depend on the presence of the dfoxo transcription factor. Whole fly transcriptome was determined with flies over-expressing a dominant negative form of the insulin receptor, or the wild-type fly, in presence or absence of dfoxo.
Project description:Transgenerational epigenetic inheritance is a subject of immense current interest. In a newly developed Drosophila model in the laboratory, genetic ablation of insulin-producing cells (IPCs) was found to affect whole -body triglyceride levels not only in the ablated flies but also in their male-line derived, non-ablated future generations. To further characterize this genetic-factor-induced transgenerational inheritance model, we have now performed whole body microarray gene expression profiling of adult males and females with genetically ablated IPCs, and of three consecutive, paternally derived non-ablated generations of adult males and females originating from ablated males. Interestingly, like altered levels of triglycerides, transcriptomic alterations are found not only in the ablated flies but also in their male-line-derived, non-ablated future generations.
Project description:To examine the effect of loss of dFOXO on the small RNA landscape of aging animals, we sequenced total small RNA collected from whole wildtype (wDah) and dFOXO-null (dfoxoΔ94) flies at young (5-6 days) and old (31 days) age.
Project description:To examine the effect of loss of dFOXO on Ago1 and Ago2 RISC loading in aging animals, we sequenced RNA immunoprecipitated from Ago1 and Ago2 RISC collected from whole wildtype (wDah) and dFOXO-null (dfoxoΔ94) flies at young (5 days) and old (35 days) age. dFOXO is a transcription factor that regulates Ago1 and Ago2 as well as lifespan.
Project description:Whole fly transcript profiles of changes upon induction of dFOXO in the gut and fat body using S1106>dFOXO flies and how this response changes when the feedback to foxo is removed in the foxo null background