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Different transcriptional strategies for zygotic genome activation in Drosophila [MNase-Seq]

ABSTRACT: During animal development, a fertilized egg is initially under the control of maternal products and only starts zygotic transcription after several cell divisions. In animals such as Xenopus, zebrafish and Drosophila, a massive increase in zygotic transcription occurs during the mid-blastula transition (MBT), when cells shift from rapid, synchronous cell divisions without gap phases to prolonged asynchronous divisions. Before MBT, only a few so-called pre-MBT genes are expressed. How transcription is set up during these early stages is poorly understood. For example, paused RNA Polymerase (Pol II) is frequently found at developmental control genes in mammalian embryonic stem cells and Drosophila embryos but when Pol II pausing is first established in the embryo is unknown. We have analyzed the genome-wide Pol II occupancy during the maternal-to-zygotic transition in hand-staged Drosophila embryos. The results show that massive Pol II recruitment and pausing is established during MBT. The ~100 genes that are transcribed before MBT are particularly short, consistent with a need for rapid transcription during these early cell divisions. Remarkably, most of these genes are transcribed without Pol II pausing and this correlates with a TATA-enriched promoter type. This suggests that distinct strategies are used for activation in the early Drosophila embryo and this may reflect general dynamic properties of promoters used throughout development. Mnase-seq in staged Drosophila embryos

ORGANISM(S): Drosophila melanogaster

SUBMITTER: Jeff Johnston

PROVIDER: E-GEOD-41686 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Different transcriptional strategies for zygotic genome activation in Drosophila [ChIP-Seq]

Project description:During animal development, a fertilized egg is initially under the control of maternal products and only starts zygotic transcription after several cell divisions. In animals such as Xenopus, zebrafish and Drosophila, a massive increase in zygotic transcription occurs during the mid-blastula transition (MBT), when cells shift from rapid, synchronous cell divisions without gap phases to prolonged asynchronous divisions. Before MBT, only a few so-called pre-MBT genes are expressed. How transcription is set up during these early stages is poorly understood. For example, paused RNA Polymerase (Pol II) is frequently found at developmental control genes in mammalian embryonic stem cells and Drosophila embryos but when Pol II pausing is first established in the embryo is unknown. We have analyzed the genome-wide Pol II occupancy during the maternal-to-zygotic transition in hand-staged Drosophila embryos. The results show that massive Pol II recruitment and pausing is established during MBT. The ~100 genes that are transcribed before MBT are particularly short, consistent with a need for rapid transcription during these early cell divisions. Remarkably, most of these genes are transcribed without Pol II pausing and this correlates with a TATA-enriched promoter type. This suggests that distinct strategies are used for activation in the early Drosophila embryo and this may reflect general dynamic properties of promoters used throughout development. ChIP-seq for Pol II, TBP, H3K4me3, H3K27me3 and H3Ac in Drosophila embryos

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ChIPseq measurement of transcriptional engagement and replication stress at the Drosophila Mid-blastula Transition

Project description:These experiments measure genome-wide RNA Pol II binding in precisely staged wild-type embryos at five time points spanning the maternal to zygotic transition (nuclear cycles 12 through 14) of Drosophila melanogaster. In addition, RNA Pol II binding at nuclear cycle 13 is measured in embryos mutant for either mei-41/ATR or zelda. To correlate RNA Pol II binding with replication stress, genome-wide profiles of Replication protein A (70kDa subunit, RpA-70 EGFP) were generated in parallel with RNA Pol II for both wild-type and zelda at nuclear cycle 13. Two replicates each for 5 time points for wild type RNA Pol II. Two replicates for mei-41 RNA Pol II, nuclear cycle 13. Two replicates for zelda RNA Pol II, nuclear cycle 13. Two replicates each for Rpa70-EGFP in wild-type or zelda, nuclear cycle 13, matched to the corresponding RNA Pol II sample.

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