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ZC3H13 and WTAP KO in hMDMs multi stimuli(NGS5421)

ABSTRACT: ZC3H13 and WTAP KO in hMDMs multi stimuli(NGS5421)

PROVIDER: PRJNA1116552 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR29181529.fastq.gz	Fastqsanger.gz
	SRR29181530.fastq.gz	Fastqsanger.gz
	SRR29181531.fastq.gz	Fastqsanger.gz
	SRR29181532.fastq.gz	Fastqsanger.gz
	SRR29181533.fastq.gz	Fastqsanger.gz

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ZC3H13 and WTAP KO in hMDMs multi stimuli(NGS5421)

Project description:Here, we revealed an immunosuppressive role of m6A mediated by ZC3H13 and WTAP on TNF-driven inflammatory phenotype in human macrophages. In the following experiment we would like to investigate if the negative regulatory role of m6A is consistent across multiple stimuli using innate ligands (LPS, R848 and polyIC). We generated ZC3H13 KOs, WTAP KOs and NTC (WT) in monocyte-derived macrophages from 4 individual human donors and stimulated cells over night with LPS (5ng/ml) , R848 (50ng/ml) , polyIC (50ng/ml) and unstimulated cells as control. This yielded a total of 48 total samples.

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NGS5523 - ATAC-seq WTAP and ZC3H13 KOs in hMDMS

Project description:human primary macropahges (hMDMs) were genetically perturbed for WTAP and ZC3H13. ATAC-seq was performed 18h post IFNg (2ng/ml) or TNFa (10ng/ml) stimulation

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NGS5523 - ATAC-seq WTAP and ZC3H13 KOs in hMDMS

Project description:NGS5523 - ATAC-seq WTAP and ZC3H13 KOs in hMDMS

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A novel RNA m6A modulator Zc3h13 plays an anchor role in facilitating nuclear RNA methylation and mouse embryonic stem cell self-renewal

Project description:N6-methyladenosine (m6A) is an abundant modification in eukaryotic mRNA, regulating mRNA dynamics by influencing mRNA stability, splicing, export and translation. Recent studies discovered m6A methyltransferases (?writer?), demethylases (?eraser?) and binding proteins (?reader?), which modulate m6A methylation. However, the precise m6A regulating machinery still remains incompletely understood. Here we demonstrate that ZC3H13, a zinc finger protein, plays an essential role in modulating m6A methylation on polyadenylated RNA in the nucleus. ZC3H13 exists in an evolutionary-conserved macromolecular complex containing WTAP, Virilizer and Hakai. We confirm the interaction among those proteins and demonstrate that knockdown of Zc3h13 in mouse embryonic stem cell (mESC) significantly decreases global m6A level on mRNA, mainly at 3? untranslated regions (3? UTR). Interestingly, fractionation assays show that upon Zc3h13 knockdown a great majority of WTAP, Virilizer and Hakai translocate to the cytoplasm and the nuclear presence of the methyltransferase Mettl3 and Mettl14 also decrease significantly. In contrast, knockdown of WTAP, Virilizer or Hakai does not change the nuclear localization of Zc3h13. This suggests that Zc3h13 is required for nuclear localization of the Zc3h13-WTAP-Virilizer-Hakai complex, which is important for RNA m6A methylation. Finally, Zc3h13 depletion, as does WTAP, Virilizer or Hakai, impairs self-renewal and triggers mESC differentiation. Taken together, our findings demonstrate that Zc3h13 plays an essential role in anchoring WTAP, Virilizer and Hakai in the nucleus to facilitate m6A methylation and to regulate mESC self-renewal.

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Binding motif analysis of METTL3，WTAP in METTL3，WTAP overexpressed Human 293T cells

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Project description:human primay macrophages (hMDMs) were genetically perturbed for METTL3 and stimulated with IFNg for 18h

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