RNA-seq analysis to assess transcriptional effects of Rp mutations in wing imaginal discs and their dependence on Xrp1
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ABSTRACT:
ORGANISM(S): Drosophila melanogaster
PROVIDER: GSE112864 | GEO | 2018/08/20
REPOSITORIES: GEO
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Project description:mRNA Seq analysis of Drosophila wing imaginal discs from Rp mutants and controls in the presence and absence of RpS12 mutations RpS12 is a regulatory gene required for Xrp1-mediated control of translation and growth in mutants for other ribosomal protein genes Loss of function mutations in Ribosomal Protein (Rp) genes are generally lethal, but heterozygous mutant phenotypes increasingly reveal previously cryptic functions of Rps in growth control and tumor suppression. Here we report that RpS12 in particular is a regulatory gene that controls its own expression levels and also signals to control hundreds of gene expression changes in mutants for haploinsufficient Rp genes, acting through the bZip protein Xrp1. Through Xrp1, RpS12 reduces the bulk translation rate of Rp mutant cells and increases the developmental rate of Rp mutant genotypes. RpS12 also has an Xrp1-independent, positive contribution to normal development, so that the effects of RpS12 mutations in different genetic backgrounds reflects the sum of the Xrp1-dependent and Xrp1-independent effects. Our findings outline a regulatory response to mutations affecting essential Rp genes that controls overall translation and growth.
2019-12-17 | GSE124924 | GEO
Project description:RNA-seq analysis to assess transcriptional effects of Rp mutations in wing imaginal discs and their dependence on Xrp1
| PRJNA449346 | ENA
Project description:BACKGROUND: Regeneration is the ability of an organism to rebuild a body part that has been damaged or amputated, and can be studied at the molecular level using model organisms. Drosophila imaginal discs, which are the larval primordia of adult cuticular structures, are capable of undergoing regenerative growth after transplantation and in vivo culture into the adult abdomen. RESULTS: Using expression profile analyses, we studied the regenerative behaviour of wing discs at 0, 24 and 72 hours after fragmentation and implantation into adult females. Based on expression level, we generated a catalogue of genes with putative role in wing disc regeneration, identifying four classes: 1) genes with differential expression within the first 24 hours; 2) genes with differential expression between 24 and 72 hours; 3) genes that changed significantly in expression levels between the two time periods; 4) genes with a sustained increase or decrease in their expression levels throughout regeneration. Among these genes, we identified members of the JNK and Notch signalling pathways and chromatin regulators. Through computational analysis, we recognized putative binding sites for transcription factors downstream of these pathways that are conserved in multiple Drosophilids, indicating a potential relationship between members of the different gene classes. Experimental data from genetic mutants provide evidence of a requirement of selected genes in wing disc regeneration. CONCLUSIONS: We have been able to distinguish various classes of genes involved in early and late steps of the regeneration process. Our data suggests the integration of signalling pathways in the promoters of regulated genes.
| S-EPMC2939566 | biostudies-literature
Project description:Regeneration is a complex process that requires a coordinated genetic response to tissue loss. Signals from dying cells are crucial to this process and are best understood in the context of regeneration following programmed cell death, like apoptosis. Conversely, regeneration following unregulated forms of death, such as necrosis, have yet to be fully explored. Here, we have developed a method to investigate regeneration following necrosis using the Drosophila wing imaginal disc. We show that necrosis stimulates regeneration at an equivalent level to that of apoptosis-mediated cell death and activates a similar response at the wound edge involving localized JNK signaling. Unexpectedly, however, necrosis also results in significant apoptosis far from the site of ablation, which we have termed necrosis-induced apoptosis (NiA). This apoptosis occurs independent of changes at the wound edge and importantly does not rely on JNK signaling. Furthermore, we find that blocking NiA limits proliferation and subsequently inhibits regeneration, suggesting that tissues damaged by necrosis can activate programmed cell death at a distance from the injury to promote regeneration.
| S-EPMC8570793 | biostudies-literature
Project description:The wing imaginal disc of Drosophila melanogaster is a prominent experimental system for research on control of cell growth, proliferation and death, as well as on pattern formation and morphogenesis during organogenesis. The precise genetic methodology applicable in this system has facilitated conceptual advances of fundamental importance for developmental biology. Experimental accessibility and versatility would gain further if long term development of wing imaginal discs could be studied also in vitro. For example, culture systems would allow live imaging with maximal temporal and spatial resolution. However, as clearly demonstrated here, standard culture methods result in a rapid cell proliferation arrest within hours of cultivation of dissected wing imaginal discs. Analysis with established markers for cells in S- and M phase, as well as with RGB cell cycle tracker, a novel reporter transgene, revealed that in vitro cultivation interferes with cell cycle progression throughout interphase and not just exclusively during G1. Moreover, quantification of EGFP expression from an inducible transgene revealed rapid adverse effects of disc culture on basic cellular functions beyond cell cycle progression. Disc transplantation experiments confirmed that these detrimental consequences do not reflect fatal damage of imaginal discs during isolation, arguing clearly for a medium insufficiency. Alternative culture media were evaluated, including hemolymph, which surrounds imaginal discs during growth in situ. But isolated larval hemolymph was found to be even less adequate than current culture media, presumably as a result of conversion processes during hemolymph isolation or disc culture. The significance of prominent growth-regulating pathways during disc culture was analyzed, as well as effects of insulin and disc co-culture with larval tissues as potential sources of endocrine factors. Based on our analyses, we developed a culture protocol that prolongs cell proliferation in cultured discs.
| S-EPMC4159298 | biostudies-literature
Project description:The apical and basolateral membranes of epithelia are insulated from each other, preventing the transfer of extracellular proteins from one side to the other. Thus, a signalling protein produced apically is not expected to reach basolateral receptors. Evidence suggests that Wingless, the main Drosophila Wnt, is secreted apically in the embryonic epidermis. However, in the wing imaginal disc epithelium, Wingless is mostly seen on the basolateral membrane where it spreads from secreting to receiving cells. Here we examine the apico-basal movement of Wingless in Wingless-producing cells of wing imaginal discs. We find that it is presented first on the apical surface before making its way to the basolateral surface, where it is released and allowed to interact with signalling receptors. We show that Wingless transcytosis involves dynamin-dependent endocytosis from the apical surface. Subsequent trafficking from early apical endosomes to the basolateral surface requires Godzilla, a member of the RNF family of membrane-anchored E3 ubiquitin ligases. Without such transport, Wingless signalling is strongly reduced in this tissue.
| S-EPMC4817240 | biostudies-literature
Project description:Screening for binding partners of the splicing factor SmD3 and changes in interaction upon depletion of the protein Ecdysoneless (Ecd) in the nubbin domain of third-instar larval wing imaginal discs.
2021-09-09 | PXD019761 | Pride
Project description:We report the changes in chromatin accessibility associated with regeneration in wing imaginal discs at two developmental timepoints during L3. Wing discs were ablated using tissue-specific expression of the pro-apoptotic gene eiger at an early timepoint (day 7) when the discs retain high regenerative capacity and at a late time point (day 9) when regenerative capacity has significantly declined. ATAC-seq was performed on these discs after 40 hr of ablation and compared to identically staged unablated discs. We detect 14,142 open chromatin peaks after merging overlapping peaks from 3 biological repeats of all 4 conditions. Of these, 349 change significantly (log2fc>0.5, p<0.1) either opening or closing upon damage in early L3 discs, while only 55 open or close in late L3 discs. We describe two consequential groups of peak changes, those that are damage responsive (DR) in early L3 (222 peaks) and those that are maturity silenced (MS), open in early damaged L3 and closed in late damaged L3 (729 peaks). 28 peaks fall in the overlap of these two groups.
2020-06-11 | GSE140755 | GEO
Project description:We report the relationship between 'absent, small or homeotic discs 2' (ASH2) occupancy, histone modifications and the transcription machinery using the wing disc as a source of chromatin. Our results indicate that genes related to development and transcriptional regulation are direct targets of ASH2 and that this protein contributes to H3K4me3 of nearby nucleosomes. Moreover, ASH2 occupancy correlates with phosphorylated forms of RNA polymerase II and histone positive marks in active genes. RNA Pol II phosphorylation in Ser5 is reduced in ash2 mutants, indicating that ASH2 plays a role in promoter stalling.
2011-02-17 | GSE24115 | GEO
Project description:The Drosophila Hedgehog receptor functions to regulate the essential downstream pathway component, Smoothened, and to limit the range of signaling by sequestering Hedgehog protein signal within imaginal disc epithelium. Hedgehog receptor function requires both Patched and Ihog activity, the latter interchangeably encoded by interference hedgehog (ihog) or brother of ihog (boi). Here we show that Patched and Ihog activity are mutually required for receptor endocytosis and degradation, triggered by Hedgehog protein binding, and causing reduced levels of Ihog/Boi proteins in a stripe of cells at the anterior/posterior compartment boundary of the wing imaginal disc. This Ihog spatial discontinuity may contribute to classically defined cell segregation and lineage restriction at the anterior/posterior wing disc compartment boundary, as suggested by our observations that Ihog activity mediates aggregation of otherwise non-adherent cultured cells and that loss of Ihog activity disrupts wing disc cell segregation, even with downstream genetic rescue of Hedgehog signal response.
| S-EPMC5668237 | biostudies-literature