Drosophila melanogaster
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA133367 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR064986.fastq.gz | Fastqsanger.gz | |||
| SRR064987.fastq.gz | Fastqsanger.gz |
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Project description:Comparison of GRO-seq with Pol II (Rpb3) ChIP-seq in Drosophila melanogaster S2 cells
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Project description:Control of RNA transcription is critical for the development and homeostasis of all organisms, and can occur at multiple steps of the transcription cycle, including RNA polymerase II (Pol II) recruitment, initiation, promoter-proximal pausing, and elongation. That Pol II accumulates on many promoters in metazoans implies that steps other than Pol II recruitment are rate-limiting and regulated 1-6. By integrating genome-wide Pol II chromatin immunoprecipition (ChIP) and Global Run-On (GRO) genomic data sets from Drosophila cells, we examined critical features of Pol II near promoters. The accumulation of promoter-proximal polymerase is widespread, occurring on 70% of active genes; and unlike elongating Pol II within the body of genes, promoter Pol II are held paused by factors like NELF, unable to transcribe unless nuclei are treated with strong detergent. Notably, we find that the vast majority of promoter-proximal Pol II detected by ChIP are paused, thereby identifying the biochemical nature of this rate-limiting step in transcription. Finally, we demonstrate that Drosophila promoters do not have the upstream divergent Pol II that is seen so broadly and prominently on mammalian promoters. We postulate this is a consequence of Drosophila’s extensive use of directional core promoter sequence elements, which contrasts with mammals’ lack of directional elements and prevalence of CpG island core promoters. In support of this idea, we show that the fraction of mammalian promoters containing a TATA box core element is dramatically depleted of upstream divergent transcription. ChIP-seq data set for Pol II (rpb3) (2 replicates).
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Project description:We performed Pol II ChIP-seq experiments with 3rd instar larva salivary glands material from male, female and MSL2 RNAi D.melanogaster, as well as from S2 cell lines material. The antibody used recognizes the Rpb3 subunit of RNA Pol II irrespective of the phosphorylation state of the Rpb1 carboxy-terminal domain, therefore capturing Pol II during all stages of transcription. Corrected processed data has been made available following Vaquerizas et al (2013). [M-^SResponse to Comments on M-^SDrosophila Dosage Compensation Involves Enhanced Pol II Recruitment to Male X-Linked PromotersM-^T, Juan M. Vaquerizas, Florence M. G. Cavalli, Thomas Conrad, Asifa Akhtar, Nicholas M. Luscombe, Science 340 (6130): 273 (2013)]
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Project description:ChIP-seq study analysing adult Drosophila melanogaster head, glial, neuronal and fat body, as well as embryonic RNA pol II and H2A.v binding by employing the GAL4-UAS system to generate GFP-fusion proteins and ChIP-seq
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Project description:We show that the non-specific lethal (NSL) complex in Drosophila binds to housekeeping genes. Individual ChIP-qPCR experiments of NSL target genes indicated a role of the NSL complex in RNA Polymerase II (Pol II) recruitment to promoters. Consequently, we obtained ChIP-Seq profiles of the Pol-II-subunit Rbp3 in S2 cells that were depleted of either NSL1 or NSL3. Rbp3-ChIP from S2 cells treated with siRNA targeted against GFP were used as a control. This experiment is related to experiment E-MTAB-214 and E-MTAB-1085.
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Project description:GRO-seq in Drosophila melanogaster S2 cells
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Project description:TF ChIP-seq Drosophila melanogaster II
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Project description:Argonaute2 and LaminB repress transcription by controlling RNA Pol II recruitment and chromatin topology in Drosophila melanogaster (ChIP-Seq)
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Project description: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.
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Project description:We use male gonads isolated from a Drosophila strain that allows us to obtain enough cells at their primitive status as the starting material to study the endogenous chromatin structure of undifferentiated cells using ChIP-seq. We integrate the ChIP-seq data with RNA-seq data that measures the transcriptome in a digital manner. Our genome-wide analyses indicate that the majority of differentiation genes in undifferentiated cells lack an active chromatin mark and paused Pol II; instead, they are associated with either the repressive H3K27me3 mark or no detectable mark. In order to address the possibility that distinct techniques are responsible for such a difference, we also use the Drosophila S2 cells to perform ChIP-seq and RNA-seq and compare the results directly with published work using ChIP-chip and microarray on S2 cells. For the S2 cell ChIP-chip data, we used data from the following paper: Muse GW, Gilchrist DA, Nechaev S, Shah R, Parker JS, Grissom SF, Zeitlinger J, Adelman K: RNA polymerase is poised for activation across the genome. /Nat Genet /2007, 39(12):1507-1511. The accession number for this data is: GSE6714. ChIP-seq: Profiling chromatin modifications using antibodies against 3 histone modifications and RNA Pol II in S2 cells Profiling chromatin structure in bam testis using antibodies against 3 histone modifications and RNA Pol II RNA-seq: Profiling transcriptome of S2 cells using RNA-seq
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