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Gene expression in AML12 cells after Uhrf2 knock-down

ABSTRACT: We identified the ubiquitin ligase Uhrf2 as a key regulator of liver regeneration. To determine the underlying mechanisms of action, we performed siRNA-mediated knockdown of Uhrf2 in AML12 cells and analysed the transcriptome of the Uhrf2-deficient cells.

ORGANISM(S): Mus musculus

PROVIDER: GSE209768 | GEO | 2023/05/25

REPOSITORIES: GEO

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BioID screen for Uhrf2 interactors in AML12 cells

Project description:We identified the ubiquitin ligase Uhrf2 as an important regulator of liver regeneration. To gain insight into the mechanisms of action of Uhrf2 in hepatocytes, we searched for Uhrf2 interactors in the AML12 hepatocyte cell line using BioID interaction screening. This resulted in the identification of major chromatin remodelling proteins as Uhrf2 interactors.

2023-06-01 | PXD041732 | Pride

UHRF2 Commissions the Accomplishment of DNA Demethylation through Allosteric Activation by 5hmC and K33-linked Ubiquitination of XRCC1

Project description:The transition of oxidized 5-methylcytosine (5mC) intermediates into base excision repair (BER) pipeline to complete DNA demethylation remains enigmatic. We report here that UHRF2, the only paralog of UHRF1 in mammals that is neither able to maintain 5mC nor to rescue Uhrf1-/- phenotype, is physically and functionally associated with BER. UHRF2 is allosterically activated by 5-hydroxymethylcytosine (5hmC) and acts as a ubiquitin E3 ligase to catalyze K33-linked polyubiquitination of XRCC1, a core subunit of the BER complex. This nonproteolytic action stimulates the interaction of XRCC1 with the ubiquitin binding domain-bearing RAD23B, leading to the formation of the TDG-RAD23B-BER complex amenable to complete DNA demethylation. Integrative epigenomic analysis in mouse embryonic stem cells reveals that the Uhrf2-coordinated TDG-RAD23B-BER complex functionally links to the completion of DNA demethylation at promoters, and that during neuronal commitment, Uhrf2-associated and H3K4me1-marked latent enhancers undergo poised-to-active transition accompanied by H3K27ac acquisition. Accordingly, Uhrf2 ablation impairs DNA demethylation and abolishes neuronal differentiation. Together, these observations highlight the essentiality of 5hmC-switched UHRF2 E3 ligase activity in commissioning the accomplishment of active DNA demethylation.

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UHRF2 regulates local 5-methylcytosine and suppresses spontaneous seizures [MeDIP-Seq, hMeDIP-Seq]

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