Drosophila transcription factors, Séance, Ouija board and Molting defective, cooperatively control ecdysone biosynthesis
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ABSTRACT:
ORGANISM(S): Drosophila melanogaster
PROVIDER: GSE104340 | GEO | 2018/02/27
REPOSITORIES: GEO
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Project description:Drosophila transcription factors, Séance, Ouija board and Molting defective, cooperatively control ecdysone biosynthesis
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Project description:Ecdysteroids are steroid hormones that control many aspects of development and physiology. During larval development, ecdysone is synthesized in an endocrine organ called the prothoracic gland through a series of ecdysteroidogenic enzymes encoded by the Halloween genes. The expression of the Halloween genes is highly restricted and dynamic, indicating that their spatiotemporal regulation is mediated by their tight transcriptional control. In this study, we report that three zinc finger-associated domain (ZAD)-C2H2 zinc finger transcription factors-Séance (Séan), Ouija board (Ouib), and Molting defective (Mld)-cooperatively control ecdysone biosynthesis in the fruit fly Drosophila melanogaster Séan and Ouib act in cooperation with Mld to positively regulate the transcription of neverland and spookier, respectively, two Halloween genes. Remarkably, loss-of-function mutations in séan, ouib, or mld can be rescued by the expression of neverland, spookier, or both, respectively. These results suggest that the three transcription factors have distinct roles in coordinating the expression of just two genes in Drosophila Given that neverland and spookier are located in constitutive heterochromatin, Séan, Ouib, and Mld represent the first example of a transcription factor subset that regulates genes located in constitutive heterochromatin.
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Project description:Samples 1-8: Tissue-specific RNA sequencing (Illumina) using dissected ring glands isolated from TWO different time points of control (phm>w1118) third instar larvae. Time points are: light phase zt0-4 (which corresponde to 2-4 hours from second to third instar larvae molt); and dark phase zt18-22 (which corresponde to 16-20 hours from second to third instar larvae molt) Samples 9-32: Tissue-specific gene expression (RNA seq Illumina) using dissected ring glands isolated from TWO different time points of third instar larvae. Genotypes were Timeless-RNAi (phm>tim-RNAi), Period-RNAi (phm>per-RNAi), UAS-TimcDNA (phm>UAS-Tim) and UAS-TimcDNA;UAS-PercDNA (phm>UAS-TimcDNA;UAS-PercDNA). Goal was to identify circadin pathway dependent gene sets in the ring gland. Time points were 2-4 hours and 18-20 hours after L2-L3 molt.
2017-07-07 | GSE83914 | GEO
Project description:The bacterial cell envelope is the frontier line of the adaptational defense that protects cells against environmental and intrinsic stressors to maintain homeostasis. Deciphering the molecular pathways involved in envelope biology is of high importance for both fundamental sciences and translational applications. Peptidoglycan (PG) is one of the main structural components of bacterial cell envelopes. It protects bacteria against mechanical and osmotic stress, confers cell shape and physical integrity. In a previous study, we identified that the Escherichia coli ElyC protein containing a highly conserved domain of an unknown function is required for envelope integrity at low temperature. ElyC was shown to be essential for PG assembly at room temperature (21°C) and to be involved in the metabolism of the lipid carrier necessary for the membrane step of the biosynthesis pathways for PG and other bacterial envelope polysaccharides. PG assembly was completely blocked in ElyC-defective cells grown at 21°C, leading to a terminal phenotype of cell lysis. The physiological importance of ElyC in bacterial cells grown at the physiological temperature of 37°C had not yet been investigated. To study this, we performed phenotypic characterization and transcriptomic profiling of ElyC-defective E. coli cells were grown at 37°C and 21°C, compared to wild-type cells. Even if ∆elyC mutant cells grow as well as wild-type cells at 37°C, microscopic analysis revealed that cellular morphology is altered by the lack of ElyC at this temperature. PG quantification and analysis confirmed that PG biosynthesis is significantly inhibited in the absence of ElyC at 37°C. We further have shown that ∆elyC mutant cells are more sensitive to PG-targeting β-lactam antibiotics than wild-type cells at 37°C. The RNA-Seq transcriptomic study showed that elimination of the ElyC evokes a highly defective cellular envelope when elyC mutant grows at 21°C, whereas at 37°C, the absence of ElyC leads to a moderate cellular response. While the PG defect of cells inactivated for ElyC is lethal at 21°C, ElyC-defective cells maintain enough PG integrity to grow and survive at 37°C. However, those cells are morphologically defective, are coping with envelope stress and are more sensitive to PG-targeting antibiotics. Overall, our data support a crucial role for ElyC in the metabolic flux for the PG biosynthesis pathway at optimal and suboptimal temperatures.
2020-03-20 | GSE141694 | GEO
Project description:Samples 1-24: Tissue-specific gene expression microarrays (Nimblegen) using dissected ring glands isolated from FOUR different time points of control (w1118, otherwise wild type) third instar larvae. Time points are 4, 8, 24 and 36 hours after the molt from second to third instar larvae. Samples 25-42: Tissue-specific gene expression microarrays (Nimblegen) using dissected ring glands isolated from TWO different time points of third instar larvae. Genotypes were phantom22-GAL4/RasV12 and phantom22/Torso-RNAi. Goal was to identify PTTH-dependent gene sets in the ring gland. Time points were 18 hours and 8 hours prior to puparium formation.
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Project description:Molting is critical for growth, development, reproduction, and survival in arthropods. Complex neuroendocrine pathways are involved in the regulation of molting and may potentially become targets of environmental endocrine disrupting chemicals (EDCs). Based on several known ED mechanisms, a wide range of pesticides has been developed to combat unwanted organisms in food production activities such as agriculture and aquaculture. Meanwhile, these chemicals may also pose hazards to nontarget species by causing molting defects, and thus potentially affecting the health of the ecosystems. The present review summarizes the available knowledge on molting-related endocrine regulation and chemically mediated disruption in arthropods (with special focus on insects and crustaceans), to identify research gaps and develop a mechanistic model for assessing environmental hazards of these compounds. Based on the review, multiple targets of EDCs in the molting processes were identified and the link between mode of action (MoA) and adverse effects characterized to inform future studies. An adverse outcome pathway (AOP) describing ecdysone receptor agonism leading to incomplete ecdysis associated mortality was developed according to the OECD guideline and subjected to weight of evidence considerations by evolved Bradford Hill Criteria. This review proposes the first invertebrate ED AOP and may serve as a knowledge foundation for future environmental studies and AOP development.
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Project description:In Drosophila melanogaster larvae the ring gland is a control center that orchestrates major developmental transitions. It is a composite organ, consisting of the prothoracic gland, the corpus allatum and the corpora cardiaca, each of which synthesizes and secretes a different hormone. Until now, the ring gland’s broader developmental roles beyond endocrine secretion have not been explored. RNA sequencing and analysis of a new transcriptome resource from D. melanogaster wandering third instar larval ring glands has provided a fascinating insight into the diversity of developmental signalling in this organ. We have found strong enrichment of expression of two gene pathways not previously associated with the ring gland: immune response and fatty acid metabolism. We have also uncovered strong expression for many uncharacterized genes. Additionally, RNA interference against ring gland-enriched cytochrome p450s Cyp6u1 and Cyp6g2 produced a lethal ecdysone deficiency and a juvenile hormone deficiency respectively, flagging a critical role for these genes in hormone synthesis. This transcriptome provides a valuable new resource for investigation of roles played by the ring gland in governing insect development.
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Project description:Ecdysteroids regulate insect growth and development through a heterodimeric complex of nuclear receptors consisting of ecdysone receptor (EcR) and ultraspiracle (USP). In the red flour beetle, Tribolium castaneum, two isoforms each of EcR and USP have been identified. Quantitative real-time reverse-transcriptase PCR (qRT-PCR) analysis showed isoform-specific developmental expression of both EcR and USP in the epidermis and the midgut dissected from the final instar larvae and pupae. Injection of double-stranded RNA (dsRNA) prepared using the common or isoform-specific regions of EcR or USP as templates caused derailment of development. EcR common region (EcRC) or EcRA dsRNA caused more severe effects, and most of the treated larvae died prior to pupation. EcRB dsRNA caused less severe effects and most of the treated larvae became pupae but showed developmental defects. Only dsRNA prepared against USP common region but not against USPA or USPB isoform-specific region caused developmental defects during larval-pupal metamorphosis. Determination of mRNA levels of EcR isoforms and 20-hydroxyecdysone-response (20E) genes (broad, E75, E74, HR3 and FTZ-F1) by qRT-PCR in the larvae injected with EcRA, EcRB or EcRC dsRNA showed that EcRA initiates ecdysteroid action by regulation the expression of EcRB and 20E-response genes. These data suggest that the EcR but not USP isoforms play distinct roles during the larval-pupal metamorphosis and EcRA plays a dominant role in transduction of ecdysteroid response in T. castaneum.
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Project description:This is part of a cross-species approach to characterize transcriptional response of the insect prothoracic gland (PG) to the prothoracicotropic hormone (PTTH). Genome-wide response of the PG to PTTH was analyzed in vitro in Bombyx and in vivo in Drosophila, and the results were compared to identify common components in the PTTH signaling pathway.
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Project description:kdm5 is an essential gene in Drosophila that has critical developmental roles in the prothoracic gland cells of the larval ring gland. We performed a bulk transcriptome analysis of the larval ring gland in w[1118] (wild type) and kdm5[140] (null mutant) in order to identify genes in the prothoracic gland involved in the lethality of kdm5 null mutants. We found that the absence of kdm5 causes dysregulation of genes involved in various metabolic pathways. In particular, genes both bound by KDM5 and differentially expressed in this cell type are involved in regulation of mitochondrial biology and autophagy.
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