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Gene expression profiling in Drosophila larvae in normal and Cmi or Trr depletion backgrounds

ABSTRACT: Gene expression profiling in Drosophila larvae in normal and Cmi or Trr depletion backgrounds

PROVIDER: PRJNA599382 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR10843344_1.fastq.gz	Fastqsanger.gz
	SRR10843344_2.fastq.gz	Fastqsanger.gz
	SRR10843345_1.fastq.gz	Fastqsanger.gz
	SRR10843345_2.fastq.gz	Fastqsanger.gz
	SRR10843346_1.fastq.gz	Fastqsanger.gz

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Gene expression profiling in Drosophila larvae in normal and Cmi or Trr depletion backgrounds

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Histone H3K4 monomethylation catalyzed by Trr and mammalian COMPASS-like proteins at enhancers is dispensable for development and viability

Project description: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.

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Stem cell proliferation is kept in check by the chromatin regulators Kismet/CHD7/CHD8 and Trr/MLL3/4

Project description:Chromatin remodeling accompanies differentiation, however, its role in self-renewal is less-well understood. We report that in Drosophila the chromatin remodeler Kismet/CHD7/CHD8 limits intestinal stem cells (ISC) number and proliferation without affecting differentiation. Stem cell-specific whole-genome profiling of Kismet revealed its enrichment at transcriptionally active regions bound by RNA Polymerase II and Brahma, its recruitment to transcription start site of activated genes and developmental enhancers and its depletion from regions bound by Polycomb, Histone H1 and heterochromatin Protein 1. We demonstrate that the Trithorax-related/MLL3/4 chromatin modifier regulates ISC proliferation, colocalizes extensively with Kismet throughout ISC genome, and co-regulates genes in ISCs, including Cbl a negative regulator of Epidermal Growth Factor Receptor (EGFR). Loss of kismet or trr leads to elevated levels of EGFR protein and signaling, thereby promoting ISC self-renewal. We propose that Kismet with Trr establishes a chromatin state that limits EGFR proliferative signaling, preventing tumor-like stem cell overgrowths.

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A small UTX stabilization domain of Trr is conserved within mammalian MLL3-4/COMPASS and is sufficient to rescue loss of viability in null animals

Project description:Catalytic-inactivating mutations within the Drosophila enhancer H3K4 monomethyltransferase Trr and its mammalian homologs, MLL3/4, cause only minor changes in gene expression compared to whole-gene deletions for these COMPASS members. To identify essential histone methylatransferase-independent functions of Trr, we screened to identify a minimal Trr domain sufficient to rescue Trr-null lethality and demonstrate that this domain binds and stabilizes Utx in vivo. Using the homologous MLL3/MLL4 human sequences, we mapped a short ~80 amino acid UTX-Stabilization-Domain (USD) that promotes UTX stability in the absence of the rest of MLL3/4. Nuclear UTX stability is enhanced when the USD is fused with the MLL4 HMG-box. Thus, COMPASS-dependent UTX stabilization is an essential non-catalytic function of Trr/MLL3/MLL4, suggesting that stabilizing UTX could be a therapeutic strategy for cancers with MLL3/4 loss-of-function mutations.

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Effect of maternal treatment on gene expression differences in Bacillus thuringiensis infected pooled larvae

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