Determination of Shavenbaby effect on histone marks in S2 cells [chipseq_histone]
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ABSTRACT: Shavenbaby (Svb) transcription factor is involved in the differenciation of epidermal cells, the homeostasis of intestinal stem cell. Svb is first synthetized as a long repressor protein called SvbRepressor (SvbREP). In presence of Pri peptides Svb Repressor is clived by the proteasoome into a shorter activator protein called Svb Activator (SvbACT). We want in this study determine the binding behaviour of the two Svb forms on S2 cells. We performed ChIPseq histones experiment in S2 cell lines which present Svb REP or SvbACT or S2 cell without Svb.
Project description:Shavenbaby (Svb) transcription factor is involved in the differenciation of epidermal cells, the homeostasis of intestinal stem cell. Svb is first synthetized as a long repressor protein called SvbRepressor (SvbREP). In presence of Pri peptides Svb Repressor is clived by the proteasoome into a shorter activator protein called Svb Activator (SvbACT). We want in this study determine the binding behaviour of the two Svb forms on S2 cells. We performed ChIPseq Svb experiment in S2 cell lines which present Svb REP or SvbACT or S2 cell without Svb.
Project description:Shavenbaby (Svb) transcription factor is involved in the differenciation of epidermal cells, the homeostasis of intestinal stem cell. Svb is first synthetized as a long repressor protein called SvbRepressor (SvbREP). In presence of Pri peptides Svb Repressor is clived by the proteasoome into a shorter activator protein called Svb Activator (SvbACT). We want in this study determine the effect of the two Svb forms on gene expression and determine the Svb target genes. We performed RNAseq experiment in S2 cell lines which present Svb REP or SvbACT or S2 cell without Svb.
Project description:Shavenbaby (Svb) transcription factor is involved in the differenciation of epidermal cells and the homeostasis of intestinal stem cells (ISCs). Svb is produced as a long repressor protein called SvbRepressor (SvbREP). In presence of Pri peptides SvbREP is processed into a shorter activator form (SvbACT) through partial proteasomal degradation of the REP domain. In ISCs, Pri triggers SvbREP to ACT maturation, while SvbREP form accumulates in differentiated enterocytes. The absence of Svb promote the apoptosis of the intestinal stem cells. In ISC, the ectopic expression of SvbREP form promote the differenciation and the SvbACT form overexpression promotes hyperplasia. (Al Hayel, et al 2020) Based on these functional analyses, Svb appears to control ISC survival and behavior, balancing proliferation and differentiation through a molecular switch between its 2 antagonistic isoforms. In this study, we determine the Svb target genes in ISCs. To this aim, we analyze the transcriptomes of ISCs upon modulation of Svb function.
Project description:Shavenbaby (Svb) transcription factor is involved in the homeostasis of intestinal stem cells (ISCs), and the differentiation of their enteroblast progeny (EB) into enterocytes (EC). Svb is produced as a long repressor protein called SvbRepressor (SvbREP). In presence of Pri peptides SvbREP is processed into a shorter activator form (SvbACT) through partial proteasomal degradation of the REP domain. In ISCs/EBs, Pri triggers SvbREP to ACT maturation, while SvbREP form accumulates in differentiated enterocytes. The absence of Svb promote the apoptosis of the intestinal stem cells. In ISCs/EBs, the ectopic expression of SvbREP form promotes the differentiation and the overexpression of SvbACT promotes hyperplasia. (Al Hayek, et al 2020) Based on these functional analyses, Svb appears to control ISC/EB survival and behavior, balancing proliferation and differentiation through a molecular switch between its 2 antagonistic isoforms. In this study, we determine the Svb target genes in different cell types along the intestinal cell lineage. This particular metadata sheet refers to enteroblast (EB) transcriptomic data (RNAseq) in control condition and upon modulation of Svb function.
Project description:Shavenbaby (Svb) transcription factor is involved in the homeostasis of intestinal stem cells (ISCs), and the differentiation of their enteroblast progeny (EB) into enterocytes (EC). Svb is produced as a long repressor protein called SvbRepressor (SvbREP). In presence of Pri peptides SvbREP is processed into a shorter activator form (SvbACT) through partial proteasomal degradation of the REP domain. In ISCs/EBs, Pri triggers SvbREP to ACT maturation, while SvbREP form accumulates in differentiated enterocytes. The absence of Svb promote the apoptosis of the intestinal stem cells. In ISCs/EBs, the ectopic expression of SvbREP form promotes the differentiation and the overexpression of SvbACT promotes hyperplasia. (Al Hayek, et al 2020) Based on these functional analyses, Svb appears to control ISC/EB survival and behavior, balancing proliferation and differentiation through a molecular switch between its 2 antagonistic isoforms. In this study, we determine the Svb target genes in different cell types along the intestinal cell lineage. This particular metadata sheet refers to early enterocytes (mEC) transcriptomic data (RNAseq) in control condition and upon modulation of Svb function.
Project description:Shavenbaby (Svb) transcription factor is involved in the homeostasis of intestinal stem cells (ISCs), and the differentiation of their enteroblast progeny (EB) into enterocytes (EC). Svb is produced as a long repressor protein called SvbRepressor (SvbREP). In presence of Pri peptides SvbREP is processed into a shorter activator form (SvbACT) through partial proteasomal degradation of the REP domain. In ISCs/EBs, Pri triggers SvbREP to ACT maturation, while SvbREP form accumulates in differentiated enterocytes. The absence of Svb promote the apoptosis of the intestinal stem cells. In ISCs/EBs, the ectopic expression of SvbREP form promotes the differentiation and the overexpression of SvbACT promotes hyperplasia. (Al Hayek, et al 2020) Based on these functional analyses, Svb appears to control ISC/EB survival and behavior, balancing proliferation and differentiation through a molecular switch between its 2 antagonistic isoforms. In this study, we determine the Svb target genes in different cell types along the intestinal cell lineage. This particular metadata sheet refers to early enterocytes (eEC) transcriptomic data (RNAseq) in control condition and upon modulation of Svb function.
Project description:The transcription factor Shavenbaby (Svb), the only member of the OvoL family in Drosophila, controls intestinal stem cell differentiation. Post-translational modification of Svb produces two protein isoforms, Svb-ACT and Svb-REP, which promote intestinal stem cell renewal or differentiation, respectively. Using engineered cell lines, we express either isoform to define their mode of action, and develop an unbiased method to identify Svb target genes in intestinal cells. Within a given cell type, Svb-ACT and Svb-REP antagonistically regulate the expression of a set of target genes, binding specific enhancers whose accessibility is constrained by. During intestinal differentiation, the set of target genes progressively changes, together with chromatin accessibility. Moreover, Svb-REP binding stabilizes three-dimensional enhancer-promoter loops, while influencing the local chromatin landscape to repress target genes. We propose that SvbACT-to-REP switch promotes enterocyte differentiation of intestinal stem cells through direct gene regulation and chromatin remodeling.
Project description:Terminal differentiation of epidermal cells in Drosophila embryos requires the activity of a transcription factor. Svb is necessary and sufficient to induce this process. pri is a regulator of Svb activity, converting it from a repressor into an activator. To characterize the downstream Svb and pri effectors in cell morphogenesis, we performed microarrays in wt, svb -/- (no gene) and pri -/- (svb repressor) mutant conditions.
Project description:Terminal differentiation of epidermal cells in Drosophila embryos requires the activity of a transcription factor. Svb is necessary and sufficient to induce this process. pri is a regulator of Svb activity, converting it from a repressor into an activator. To characterize the downstream Svb and pri effectors in cell morphogenesis, we performed microarrays in wt, svb -/- (no gene) and pri -/- (svb repressor) mutant conditions. Embryos were selected at a precise 13-15h (after egg laying) stage of development, and manually genotyped for each condition: wt (W samples), svb -/- (R samples) and pri -/- (P samples). 5 independent replicates of 200 embryos are used for each condition. RNA were extracted and hybridized on Affymetrix microarrays.