A genome-wide survey for a Juvenile hormone response element, DR4, in the fruit fly, Drosophila melanogaster
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ABSTRACT:
ORGANISM(S): Drosophila melanogaster
PROVIDER: GSE6999 | GEO | 2009/12/31
SECONDARY ACCESSION(S): PRJNA99279
REPOSITORIES: GEO
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Project description:Microarray analyses identified 713 genes whose expression increased or decrease by 1.5-fold with a P value <0.01 in response to exposure to JH for 3-12 hr. 497 were up-regulated, and 228 were down-regulated. 12 genes showed differentially regulated pattern that depended on the time of exposure to JH. 73 of these genes (57 genes up-regulated and 17 genes down-regulated) showed the presence of DR4 element in their promoter regions. We also screened the promoter regions of genes whose products are distributed in different cellular locations and various functional groups and found that the DR4 element is present in 275 out of 2850 genes screened. The distribution of DR4 element containing genes varied depending on both cellular location and gene function. The genes whose products are localized to the nucleus or cytoplasm showed higher proportion of DR4 element containing genes when compared to the genes coding for products localized to the membrane. Genes belonging to immune response and ligand activated receptor functional groups showed higher number of genes that contained DR4 elements when compared to the genes belonging to protein kinase or transmembrane receptor functional groups. The DR4 elements identified in the promoter regions of D. melanogaster genes bound to the nuclear proteins isolated from JH III-treated Drosophila L57 cells. In addition, the expression of DR4 element containing genes identified from various functional groups was induced by JH III in Drosophila mbn2 cells grown in the medium containing 1 µM JH III. Presence of DR4 element in JH-responsive genes is conserved across species. This study also demonstrates the feasibility of using genome-wide analysis for identification of genes regulated by various hormones and other signaling molecules. Keywords: Time course We performed microarray analysis using RNA isolated form L57 cells that were exposed to 1 µM JH III for 0, 3, 6 and 12 hr and identified 497 genes that are induced by JH and 228 genes that are down-regulated by JH. Then, we searched the promoter regions of these genes and found that 57 out of these 497 JH induced genes, and 17 out of these 228 JH down-regulated genes had conserved DR4 element present in their promoters.
2009-12-31 | E-GEOD-6999 | biostudies-arrayexpress
Project description:Juvenile hormone (JH) and 20-hydroxy-ecdysone (20E) are highly versatile hormones, coordinating development, growth, and reproduction in insects. Pulses of 20E provide key signals for initiating developmental and physiological transitions, while JH promotes or inhibits these signals in a stage-specific manner. Previous evidence suggests that JH and 20E might modulate innate immunity, but whether and how these hormones interact to regulate the immune response remains unclear. Here we show that JH and 20E have antagonistic effects on the expression of antimicrobial peptides (AMPs) in Drosophila melanogaster. In S2* cells challenged with bacterial peptidoglycans, 20E induces promoter activity and expression of AMPs in a dose-dependent manner, while JH III and its synthetic analogs (JHa) methoprene and pyriproxyfen abolish this 20E-dependent response. Using microarrays and GFP reporter gene assays in adult flies, we confirm that JH is a hormonal immuno-suppressor in vivo. When silencing both partners of the ecdysone receptor (EcR ) / ultraspiracle (USP) heterodimer with RNAi in S2* cells, 20E fails to activate Diptericin (Dpt) expression, suggesting that 20E regulates expression of this gene through EcR / USP signaling. In contrast, silencing methoprene-tolerant (MET), a candidate JH receptor, does not impair the immuno-suppressive action of JH III and JHa, indicating that in this context MET does not function as a JH receptor. Our results suggest that the balance of 20E and JH is a major determinant of immune homeostasis in insects. Experiment Overall Design: To examine the transcriptional response of y, w flies to treatment with exogenous JH, we performed a microarray experiment on females treated with JH or solvent (control). Flies were grown on regular yeast diet, switched to no-yeast food within one hour of eclosion, and yeast-starved for 5 days posteclosion to lower their endogenous JH titer and to synchronize their physiology (cf. Tu and Tatar, 2003; Gershman et al., 2007). Subsequently, we anesthetized flies on ice and topically treated them with 0.1 ïl of 187 mM JH III in acetone or with 0.1ï ïl 100% acetone (control) using a 1 ïl Hamilton syringe with a repeating dispenser. 12 hours after hormone administration, samples were snap-frozen in liquid nitrogen and stored at -80°C. RNA was isolated from samples (2 JH samples, 2 control samples, each with 30 females) by lysis, as described in Gershman et al. (2007). cDNA products were hybridized at the Brown University Genomics Core Facility to Affymetrix GeneChip Drosophila_1 Genome Arrays (2 replicate chips per treatment). The dataset consisted of 14,009 probe sets, with 6,142 probe sets annotated. Expression data were analyzed for significant over- or underrepresentation of gene ontology (GO) terms with the web application FatiGO (Al-Shahrour et al., 2004), using a two-fold change criterion. To test whether JH treatment significantly suppresses expression of AMPs we used t-tests implemented in JMP IN 5.1.
2008-06-16 | E-GEOD-9001 | biostudies-arrayexpress
Project description:Adult reproductive diapause is a powerful overwintering strategy for many continental insect species including bumblebees, which enables queens to survive several months through harsh winter conditions and then build new beehives in the following spring. There are few reports regarding the molecular regulatory mechanism of reproductive diapause in Bombus terrestris, which is an important pollinators of wild plants and crops, and our previous researches identified the conditions for reproductive diapause of year-round mass rearing. Here, we performed combined RNA sequencing transcriptomics and quantitative proteomic analyses in different development phases relate to reproductive diapause. According to the overall analysis, we found these differentially expressed proteins/genes act in the citrate cycle, insect hormone biosynthesis, insulin and mTOR signalling pathway. To get better sense of the reproductive diapause regulated mechanism, some genes regulated JH synthesis, insulin/ TOR signal pathway were detected, the BtRheb, BtTOR, BtVg and BtJHAMT had lower expression levels in diapause queens, and the JH III titers levels and some metabolic enzymes activities were significantly up-regulated in found post-diapause queens. After microinjected insulin-like peptides (ILPs) and JH analog (JHA), some indicators shows the significantly changes of hormones, cold tolerance substances, metabolic enzymes and reproduction. Along with other related researches, a reproductive diapause regulated model during B. terrestris year-round mass rearing process was establishment. This study contribute to a comprehensive view and the molecular regulate mechanism of productive diapause in eusocial insect.
2021-04-06 | PXD017616 | Pride
Project description:In honeybees, differential feeding of female larvae promotes the occurrence of two different phenotypes, a queen and a worker, from identical genotypes. Nutritional factors affect both endocrine responses and gene expression and, thus, change the larval developmental pattern. It is widely accepted that two types of alterations occur during this process: incremental alterations, which affect general growth, and character state alterations that result in the presence or absence of specific structures. Previous studies revealed a link between incremental alterations and differential expression of physiometabolic genes. Here, we report evidence for molecular changes that affect character states, controlling the expression of worker-like characteristics. By using cDNA microarray analyses of more than 6,000 Apis mellifera ESTs we found 240 differentially expressed genes (DEG) between developing queens and workers. Comparative analyses showed that many genes recorded as up-regulated in prospective workers appear to be unique to A. mellifera suggesting that the workers developmental pathway involves the participation of novel genes. We also show that workers up-regulate more developmental genes than queens, whereas queens up-regulate a greater predominance of physiometabolic genes. The latter include genes coding for metabolic enzymes and genes whose products are known to regulate the rate of mass-transforming processes and the general growth of the organism (e.g., tor). Many DEG are likely to be involved in processes favoring the development of caste-specific structures, like brain, legs and ovaries, as well as genes that code for cytoskeleton constituents. Treatment of developing worker larvae with juvenile hormone (JH) revealed 52 JH responsive genes, specifically during the critical period of castes development. Using Gibbs sampling and Expectation Maximization algorithms, we discovered eight overrepresented cis-elements from 4 gene groups. Graph theory and complex networks concepts were adopted to attain powerful graphical representations of the interrelation between cis-elements and genes and objectively quantify the degree of relationship between these entities. We suggest that clusters of functionally related DEGs are co-regulated during caste development in honeybees. This network of interactions is activated by nutrition-driven stimuli in early larval stages. Our data are consistent with the hypothesis that JH is a key component of the developmental determination of queen-like characters. We propose a conceptual model of caste differentiation in A. mellifera based on gene-regulatory networks. Keywords: caste differentiation during larval development; JH effect
2007-05-31 | GSE6452 | GEO
Project description:Juvenile hormone (JH) and 20-hydroxy-ecdysone (20E) are highly versatile hormones, coordinating development, growth, and reproduction in insects. Pulses of 20E provide key signals for initiating developmental and physiological transitions, while JH promotes or inhibits these signals in a stage-specific manner. Previous evidence suggests that JH and 20E might modulate innate immunity, but whether and how these hormones interact to regulate the immune response remains unclear. Here we show that JH and 20E have antagonistic effects on the expression of antimicrobial peptides (AMPs) in Drosophila melanogaster. In S2* cells challenged with bacterial peptidoglycans, 20E induces promoter activity and expression of AMPs in a dose-dependent manner, while JH III and its synthetic analogs (JHa) methoprene and pyriproxyfen abolish this 20E-dependent response. Using microarrays and GFP reporter gene assays in adult flies, we confirm that JH is a hormonal immuno-suppressor in vivo. When silencing both partners of the ecdysone receptor (EcR ) / ultraspiracle (USP) heterodimer with RNAi in S2* cells, 20E fails to activate Diptericin (Dpt) expression, suggesting that 20E regulates expression of this gene through EcR / USP signaling. In contrast, silencing methoprene-tolerant (MET), a candidate JH receptor, does not impair the immuno-suppressive action of JH III and JHa, indicating that in this context MET does not function as a JH receptor. Our results suggest that the balance of 20E and JH is a major determinant of immune homeostasis in insects. Keywords: Topical hormone treatment of whole flies
2007-12-31 | GSE9001 | GEO
Project description:The freshwater zooplankton Daphnia magna has been used for the validation of chemical toxicity tests such as OECD Test Guideline 202 and 211. Previously, it has been demonstrated that treatment of juvenile hormones (JHs) or their analogs to mother daphnids could induce male offspring production. Based on this funding, a rapid screening method for detection of chemical with JH-activity was recently developed by using adult D. magna. This screening system can validate whether arbitrary chemical has JH-activity by investigating the male offspring inducibility. Although this short-term screening system will be possible to enhance a throughput of detection efficiency, JH-responsive genes in the ovary including developing oocytes at JH-sensitive period for male sex determination are still largely undescribed. Here, we conducted comparative microarray analyses using ovary treated by fenoxycarb (Fx; artificial JH agonist) and methyl farnesoate (MF; a putative innate JH in daphnids) to make the comprehensive gene catalogs in response to JH molecules in ovary including developing oocytes at JH-sensitive period for male sex determination. We successfully created the comprehensive ovarian gene catalogs in response to Fx and MF. Furthermore, we demonstrated that the hemoglobin genes are well-conserved JH-responsive element even in the ovary, and a potential toxic effect of JH agonist is suppression of vitellogenin gene expression that might cause reduction of offspring number. This is first report demonstrating the differential transcriptomics profiles between MF and artificial JH agonist in D. magna ovary, making it possible for understanding the tissue-specific mode-of-action of JH molecule.
2017-05-01 | GSE81083 | GEO
Project description:Juvenile hormone (JH) is produced and secreted by corpora allata (CA) during larval stages. JH biosynthesis from acetyl-CoA includes 13 enzymatic steps. Genes for 10 of those enzymes have been identified from Bombyx mori. It was also reported that they are basically expressed selectively in CA. Genes for the other 3 enzymes were recently reported from other insects, i.e., farnesyl diphosphate pyrophosphatase (FPPP) from Drosophila and farnesol dehydrogenase (FolD) and farnesal dehydrogenase (FalD) from Aedes. In order to reveal the expression profiles of these genes in Bombyx CA and to find other candidate genes involved in JH biosynthesis, we performed a transcriptomic analysis in corpora cardiaca-CA (CC-CA) complexes using a custom-made DNA microarray on which sequences from CC-CA EST libraries and genome-widely predicted genes are loaded. CC-CA complexes vigorously expressed JH-biosynthetic genes identified in Bombyx as well as genes involved in methyl-group metabolism and ribosomal protein genes; this is suitable for JH production. Bombyx counterparts of Drosophila FPPP and Aedes FolD genes, however, showed quite low level of expression. On the other hand, some other genes probably encoding oxidoreductases were expressed highly and selectively in CC-CA complex. Their products might function as FolD and/or FalD in Bombyx.
2014-12-31 | GSE41805 | GEO
Project description:Bioinsecticides and transgenic crops based on the bacterial pathogen Bacillus thuringiensis (Bt) can effectively control diverse agricultural insect pests, nevertheless, insect resistance evolution without causing obvious fitness costs has seriously eroded the sustainable use of these Bt products. Recently, we have discovered that an increased titer of juvenile hormone (JH) favors an insect host (Plutella xylostella) to enhance fitness whilst resisting the Bt pathogen, however, the underlying regulatory mechanisms of the increased JH titer is obscure. Here, we define the involvement of N6-methyladenosine (m6A) RNA modification in modulating the availability of JH in this process. Specifically, we found that two m6A methyltransferase subunit genes, PxMettl3 and PxMettl14, repressed the expression of a key JH-degrading enzyme JH esterase (JHE) to induce an increased JH titer, mitigating the fitness costs associated with a robust defense against the Bt pathogen. This study identifies an as-yet uncharacterized m6A-mediated epigenetic regulator of insect hormones for maintaining fitness during pathogen defense and unveils an emerging Bt resistance-related m6A methylation atlas in insects, which further expands the functional landscape of m6A modification and showcases the pivotal role of epigenetic regulation in host-pathogen interactions.
2023-11-10 | GSE247107 | GEO
Project description:Juvenile hormone (JH) serves vital roles in insect reproduction, development, and many aspects of physiology. JH primarily acts at the gene-regulatory level through an intracellular receptor. The JH receptor (JHR) is a ligand-activated complex of transcription factors of the bHLH-PAS family, consisting of the JH-binding protein Methoprene-tolerant (MET) and its partner Taiman (TAI). Initial studies have indicated significance of post-transcriptional modification (phosphorylation), subunit assembly, and nucleocytoplasmic transport of JHR in JH signaling. However, our knowledge of JHR regulation at the protein level remains rudimentary, partly due to the difficulty of obtaining functional JHR proteins in a purified form. Here we present successful fermentation-level purification of JHR complexes of MET and TAI proteins from two insect species, the beetle Tribolium castaneum and the mosquito Aedes aegypti. The recombinant JHR subunits from each species were co-expressed using a baculovirus system in an insect cell line and purified through affinity steps, yielding soluble proteins, capable of binding both the hormonal ligand (JH III) and DNA bearing cognate JH-response elements. The present quantitative phosphoproteomcis analysis uncovered multiple phosphorylation sites in the TcMET protein, some of which were induced by methoprene. A functional bipartite nuclear localization signal, straddled by phosphorylated residues, was found within the disordered C-terminal region of TcMET. Our present characterization of the recombinant JHR is a primary step towards understanding JHR regulation.
2022-02-16 | PXD028599 | Pride
Project description:Juvenile hormone (JH) serves vital roles in insect reproduction, development, and many aspects of physiology. JH primarily acts at the gene-regulatory level through an intracellular receptor. The JH receptor (JHR) is a ligand-activated complex of transcription factors of the bHLH-PAS family, consisting of the JH-binding protein Methoprene-tolerant (MET) and its partner Taiman (TAI). Initial studies have indicated significance of post-transcriptional modification (phosphorylation), subunit assembly, and nucleocytoplasmic transport of JHR in JH signaling. However, our knowledge of JHR regulation at the protein level remains rudimentary, partly due to the difficulty of obtaining functional JHR proteins in a purified form. Here we present successful fermentation-level purification of JHR complexes of MET and TAI proteins from two insect species, the beetle Tribolium castaneum and the mosquito Aedes aegypti. The recombinant JHR subunits from each species were co-expressed using a baculovirus system in an insect cell line and purified through affinity steps, yielding soluble proteins, capable of binding both the hormonal ligand (JH III) and DNA bearing cognate JH-response elements. The present shotgun LC-MSMS analyses identified at least ten potential phosphoserine and phosphothreonine residues located in the C-terminal disordered region of TcMET. A functional bipartite nuclear localization signal, straddled by some of these phosphorylated residues, was subsequently identified within the disordered C-terminal region of TcMET.
2022-02-16 | PXD028394 | Pride