Expression profiling by high throughput sequencing Genome binding/occupancy profiling by high throughput sequencing
Summary
Histone H3 lysine 4 monomethylation (H3K4me1) is an evolutionarily conserved feature of enhancer chromatin catalyzed by the COMPASS-like methyltransferase family that includes Trr in Drosophila melanogaster and MLL3 (encoded by KMT2C) and MLL4 (encoded by KMT2D) in mammals. Here we demonstrate that Drosophila embryos expressing catalytically deficient Trr eclose and develop to productive adulthood. Parallel experiments with a trr allele that augments enzyme product specificity show that conversion of H3K4me1 at enhancers to H3K4me2 and H3K4me3 is also compatible with life and results in minimal changes in gene expression. Similarly, loss of the catalytic SET domains of MLL3 and MLL4 in mouse embryonic stem cells (mESCs) does not disrupt selfrenewal. Drosophila embryos with trr alleles encoding catalytic mutants manifest subtle developmental abnormalities when subjected to temperature stress or altered cohesin levels. Collectively, our findings suggest that animal development can occur in the context of Trr or mammalian COMPASS-like proteins deficient in H3K4 monomethylation activity and point to a possible role for H3K4me1 on cis-regulatory elements in specific settings to fine-tune transcriptional regulation in response to environmental stress.
Overall design
ChIP-seq and RNA-seq experiments were conducted in Drosophila wing imaginal discs and adult brain tissues from transgenic flies expressing either a catalytic-deficient (Trr-C2398A) or catalytic-hyperactive (Trr-Y2383F) Trr enzyme. ChIP-seq and RNA-seq were also conducted in mouse embryonic stem cells in which CRISPR-Cas9 was used to delete the SET domain of both MLL3 and MLL4 (MLL3/4-delSET).
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