Expression profiling by high throughput sequencing Genome binding/occupancy profiling by high throughput sequencing Other
Summary
Before zygotic genome activation (ZGA) the quiescent genome undergoes reprogramming to transition into the transcriptionally active state. However, the mechanisms underlying euchromatin establishment during early embryogenesis remain poorly understood. Here, we show that histone H4 lysine-16 acetylation (H4K16ac) is maintained from oocytes to fertilized embryos in Drosophila and mammals. H4K16ac forms large domains that control nucleosome accessibility of promoters prior to ZGA in flies. Maternal depletion of MOF acetyl-transferase leading to H4K16ac loss causes aberrant Pol II recruitment, compromises the 3D organization of the active genomic compartments during ZGA and consequently causes downregulation of post-zygotically expressed genes. Germline depletion of histone deacetylases revealed that other acetyl marks cannot compensate for H4K16ac loss in the oocyte. Moreover, zygotic re-expression of MOF was neither able to restore embryonic viability nor onset of X chromosome dosage compensation. Thus, maternal H4K16ac provides an instructive function to the offspring, priming future gene activation.
Overall design
DNA: We have generated a) Mnase ChIPseq profiles of H4K16ac from hand staged Drosophila embryos at various developmental stages derived from WT and MSL complex RNAi conditions b) ATAC-seq of embryo nc9-13 and nc14 upon maternal depletion of control mof RNAi or msl-3 and c) Hi-C of embryos upon control of mof maternal depletion at st5 (nc14) and st15. RNA: We have generated poly-A RNA-seq datasets derived from: a) Single embryo mRNA seq of st5 and st7 embryos after maternal mof depletion. The gender of embryos was identified post-sequencing using expression pattern of msl-2, sxl, tra and tra2. b) St15 embryo male and female separated embryos after maternal mof depletion. c) RNA-seq from mof depleted S2 cells.
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