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Ecd protein interactome in Drosophila melanogaster S2 cells

ABSTRACT: Identification of the interaction partners of the protein ecdysoneless (Ecd) in Drosophila melanogaster S2 cells as well as profiling of the changes in binding for mutant, truncated Ecd del34 protein.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Drosophila Melanogaster Drosophila Melanogaster (fruit Fly)

SUBMITTER: Steffen Erkelenz

LAB HEAD: Mirka Uhlirova

PROVIDER: PXD019838 | Pride | 2021-09-09

REPOSITORIES: Pride

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Ecd promotes U5 snRNP maturation and Prp8 stability.

Erkelenz Steffen S Stanković Dimitrije D Mundorf Juliane J Bresser Tina T Claudius Ann-Katrin AK Boehm Volker V Gehring Niels H NH Uhlirova Mirka M

Nucleic acids research 20210201 3

Pre-mRNA splicing catalyzed by the spliceosome represents a critical step in the regulation of gene expression contributing to transcriptome and proteome diversity. The spliceosome consists of five small nuclear ribonucleoprotein particles (snRNPs), the biogenesis of which remains only partially understood. Here we define the evolutionarily conserved protein Ecdysoneless (Ecd) as a critical regulator of U5 snRNP assembly and Prp8 stability. Combining Drosophila genetics with proteomic approaches ...[more]

PMID: 33444449

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SmD3 protein interactome in Drosophila melanogaster wing imaginal discs

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

Pancreatic Cancer HPAF/CD18 cells: Ecd knockdown vs Scrambled

Project description:Purpose: To study the expression and function of a novel cell cycle regulatory protein, human ecdysoneless (Ecd), during pancreatic cancer (PC) pathogenesis. Experimental Design: Immunohistochemical expression profiling of Ecd was done in non-neoplastic normal pancreatic tissues and pancreatic ductal adenocarcinoma lesions (from tissue microarray and Rapid Autopsy program) as well as precancerous PanIN lesions and metastatic organs. To analyze the biological significance of Ecd in PC progression, Ecd was stably knocked down in PC cell line followed by in vitro and in vivo functional assays. Results: Normal pancreatic ducts show very weak to no Ecd expression compared to significant positive expression in PC tissues (mean±SE composite score: 0.3±0.2 and 3.8±0.2 respectively, p<0.0001) as well as in PanIN precursor lesions with a progressive increase in Ecd expression with increasing dysplasia (PanIN-1 to PanIN-3). Analysis of matched primary tumors and metastases from PC patients revealed that Ecd is highly expressed in both primary pancreatic tumor and in distant metastatic sites. Further, knockdown of Ecd suppressed cell proliferation in vitro and tumorigenicity of PC cells in mice orthotopic tumors. Microarray study revealed that Ecd regulates expression of glucose transporter GLUT4 in PC cells and was subsequently shown to modulate glucose uptake, lactate production and ATP generation by PC cells. Finally, knockdown of Ecd also reduced level of pAkt, key signaling molecule known to regulate aerobic glycolysis in cancer cells. Conclusion: Ecd is a novel tumor promoting factor that is differentially expressed in pancreatic cancer and potentially regulates glucose metabolism within cancer cells. Two-condition experiment, Ecd knockdown vs Scrambled cells. Biological replicates: 3 Ecd knockdownl, 3 Scrambled, independently grown and harvested. One replicate per array

2012-08-09 | E-GEOD-39988 | biostudies-arrayexpress

Characterization of the redox proteome of Drosophila melanogaster S2 cells and w1118 fly eyes.

Project description:Reactive oxygen species (ROS) serve as intracellular signaling molecules; however, in excess ROS molecules are damaging to biomacromolecules such as DNA, lipids, and proteins. The excess accumulation of ROS leads to oxidative stress, disrupting redox homeostasis in the cell and has been linked with aging and neurodegenerative disease. The eye is particularly vulnerable to oxidative stress due to the tissue’s high energy demand and generation of ROS by products. In proteins, the thiol group on cysteine residues is susceptible to oxidation. Cysteine residues have multiple oxidation states that can be classified into two categories—reversible and irreversible. Irreversible oxidation states include sulfinic and sulfonic acids, which may alter or impair the activity of the protein depending on the position of the cysteine residue. In contrast, reversible oxidation states include sulfenic acid or inter- and intramolecular disulfides, and can often function as redox sensors in the cell. Here, we sought to evaluate how increasing amounts of oxidative stress alters the redox proteome landscape in Drosophila. To characterize the redox proteome, we developed an iodoTMT-multiplex isolation, labeling, and enrichment method to identify cysteine availability and potential oxidation in both S2 cells and w1118 fly eyes. To do this, we exposed S2 cells to 20mM H2O2 relative to control (untreated), and w1118 flies to prolonged blue light versus an untreated control, followed by redox proteome profiling with iodoTMT-sixplex reagents.

2023-04-27 | PXD034050 | Pride

RNA-Seq in Drosophila melanogaster S2 cells

Project description:RNA-Seq procedure was used for analysis of expression in S2 cells.

2020-12-31 | GSE145320 | GEO

Drosophila melanogaster S2-GMR cells, second biological replicate

Project description:High-throughput sequencing of Drosophila melanogaster small RNAs from S2 cells. total RNA, ~18-26nt RNAs isolated using PAGE, ligation to adapters requires 5' monophosphate and 3' OH For data usage terms and conditions, please refer to http://www.genome.gov/27528022 and http://www.genome.gov/Pages/Research/ENCODE/ENCODEDataReleasePolicyFinal2008.pdf

2009-01-22 | GSE14488 | GEO

ChIP-seq of E2f, Max, Myb and Myc in Drosophila melanogaster S2 cells for mapping their genome-wide binding sites

Project description:ChIP-seq for the strongest cell cycle regulator transcription factors in Drosophila Melanogaster S2 cells. These assays have been used to validate the direct transcriptional targets of the same transcription factors investigated in RNA-seq (E-MTAB-1364) and Affymetrix microarray experiments (E-MTAB-453). ChIP-seq assays have been done with tagged fusion proteins (for example, since we dont have functional E2f antibodies against endogenous E2f , we are transfecting v5-tagged-E2f-ORF to S2 cells and then use antibodies against v5 to detect the signal from E2f binding). If the ChIP-seq has been done with tagged fusion proteins (such as v5-tagged-E2f-ORF), the protein expression has been induced with CuSO4 treatment 48h prior to cell crosslinking & lysis. Our fusion protein constructs are driven by metallothionein promoter, which is induced by CuSO4. E-MTAB-1648, E-MTAB-1364 and E-MTAB-453 are all data from: Bonke M, et al. (2013) Transcriptional networks controlling the cell cycle. G3 (Bethesda) 3, 75-90, PMID: 23316440.

2013-08-01 | E-MTAB-1648 | biostudies-arrayexpress

Genome wide distribution of dH1K27me2 in Wild Type Drosophila melanogaster S2 cells

Project description:Genome wide distribution of dH1K27me2 in Wild Type Drosophila melanogaster S2 cells

2022-08-02 | GSE167018 | GEO

An analysis of the impact of infection by Buchnera aphidicola APS on gene expression of Drosophila S2 cells.

Project description:An analysis of the impact of infection by Buchnera aphidicola APS (isolated from Acyrthosiphon pisum strain LL01) on gene expression of S2 cells. All comparisons are made against a pool of RNA from S2 cells not exposed to B. aphidicola. B. aphidicola freshly isolated from the aphids, and data are collected at 1, 6 and 24 hours after exposure of S2 cells to the B. aphidicola preparation. Keywords: time course, Buchnera aphidicola APS, aphids, Drosophila melanogaster S2 cells All comparisons are made against a pool of RNA from S2 cells not exposed to B. aphidicola. B. aphidicola freshly isolated from the aphids, and data are collected at 1, 6 and 24 hours after exposure of S2 cells to the B. aphidicola preparation.

2008-12-14 | E-GEOD-11012 | biostudies-arrayexpress

ChIP-seq of GFP-tagged dCoREST, dLSD1, dL(3)mbt and dG9a in D. melanogaster S2 cell lines

Project description:We employed CRISPR/Cas-mediated genome editing to generate S2 cell lines expressing GFP-tagged dCoREST, dL(3)mbt, dLSD1 and dG9a, respectively. This allowed us to determine the genome-wide binding profiles for these proteins by ChIP-seq using the same antibody (anti-GFP) in each case.

2019-11-04 | E-MTAB-8341 | biostudies-arrayexpress

Project description:S2 Metagenome data

| PRJNA790701 | ENA

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