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Enrichment of dsRNA in primary intestinal epithelium cells with Kat2a/Kat2b DKO.

ABSTRACT: Enrichment of dsRNA in primary intestinal epithelium cells with Kat2a/Kat2b DKO.

PROVIDER: PRJNA987638 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR25033428_1.fastq.gz	Fastqsanger.gz
	SRR25033428_2.fastq.gz	Fastqsanger.gz
	SRR25033429_1.fastq.gz	Fastqsanger.gz
	SRR25033429_2.fastq.gz	Fastqsanger.gz
	SRR25033430_1.fastq.gz	Fastqsanger.gz

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Enrichment of dsRNA in primary intestinal epithelium cells with Kat2a/Kat2b DKO.

Project description:dsRIP-seq was conducted to determine differential enrichment of dsRNA in whole intestinal epithelial cells between Kat2a/Kat2b DKO and controls.

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Transcriptomic changes in primary intestinal epithelium cells with Kat2a/Kat2b DKO.

Project description:RNA-seq was conducted to determine differentially expressed genes in whole intestinal epithelial cells between Kat2a/Kat2b DKO and controls.

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Transcriptomic changes in primary intestinal epithelium cells with Kat2a/Kat2b DKO.

Project description:Transcriptomic changes in primary intestinal epithelium cells with Kat2a/Kat2b DKO.

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Divergent Functions of Histone Acetyltransferase Paralogs KAT2A and KAT2B in Keratinocyte Stemness and Differentiation

Project description:The mammalian epidermis undergoes constant renewal replenished by a pool of stem cells and terminal differentiation of their progeny. This is accompanied by changes in gene expression and morphology orchestrated, in part, by epigenetic modifiers. Here, we defined the role of histone acetyltransferase KAT2A in epidermal homeostasis and provided a comparative analysis that revealed key functional divergence with its paralogue, KAT2B. In contrast to KAT2B's reported function in epidermal differentiation, KAT2A supports the undifferentiated state in keratinocytes. RNA-seq analysis of KAT2A- and KAT2B- depleted keratinocytes revealed dysregulated epidermal differentiation. Depletion of KAT2A led to premature expression of epidermal differentiation genes in the absence of inductive signals, whilst loss of KAT2B delayed differentiation. KAT2A acetyltransferase activity was indispensable in regulating epidermal differentiation gene expression. The metazoan-specific N-terminus of KAT2A was also required to support its function in keratinocytes. We further showed that the interplay between KAT2A- and KAT2B- mediated regulation was important for normal cutaneous wound healing in vivo. Overall, these findings reveal a distinct mechanism in which keratinocytes utilize a pair of highly homologous histone acetyltransferases to support divergent functions in self-renewal and differentiation processes.

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Human KAT2A/B-acetylome MudPIT

Project description:Comprehensive identification of acetylated proteins targeted by the lysine acetyltransferases KAT2A (GCN5) and KAT2B (PCAF), in human cells. KAT2A/B-dependent acetylated proteins were identified by comparing acetylated proteins in whole cell extract of control cells and cells in which KAT2A and KAT2B were simultaneously knocked down. The identification of acetylated proteins was performed using an enrichment-free approach. In order to do so, while facing the challenge of identifying low abundant acetylated peptides within a complex sample mixture, we have screened acetylated peptides starting from a large amount of material, equivalent to 50ug of protein digest (maximum loading capacity of our analytical chromatographic column). By analysing such a large amount of peptides, the level of acetylated peptides was then increased and thus acetylated peptides became easily detectable by mass spectrometry, without the need for enrichment.

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KAT2A and KAT2B prevents double-stranded RNA formation and interferon signaling to maintain intestinal stem cell renewal

Project description:Histone acetyltransferases KAT2A and KAT2B are paralogs highly expressed in the intestinal epithelium, but their functions are not well understood. In this study, double knockout of murine Kat2 genes in the intestinal epithelium was lethal, resulting in diminished H3K9ac expression, loss of stem cells, and robust activation of interferon signaling. Use of pharmacological agents and sterile organoid cultures indicated a cell-intrinsic double-stranded RNA trigger for interferon signaling. Acetyl-proteomics and dsRIP-seq were employed to interrogate the mechanism behind this response, which identified self-derived, mitochondria- encoded double-stranded RNA as the source of intrinsic interferon signaling. KAT2A and KAT2B therefore play an essential role in regulating mitochondrial functions as well as maintaining intestinal health.

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Kat2a regulates hippocampal memory formation [RNA-seq]

Project description:We sequenced mRNA from 24 samples extracted from mouse CA1 tissue to generate the first CA1-specific murine transcriptome and the first CA1-transcriptome in response to environmental novelty under normal and Kat2a-loss-of-function conditions. Samples were divded in 4 groups: A: Control naM-CM-/ve (n=6), B: control novelty-exposed (n=5), C: Kat2a cKO naM-CM-/ve (n=6), D: Kat2a cKO novelty-exposed (n=7). Pairwise comparisons for AvsB, AvsC, BvsD and CvsD were performed using DESeq2.

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Project description:KAT2A and KAT2B prevent double-stranded RNA accumulation and interferon signaling to maintain intestinal stem cell renewal

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