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Clinical and molecular significance of the RNA m6A methyltransferase complex in prostate cancer

ABSTRACT: shRNAs targeting METTL3 used to deplete METTL3 in castrate-resistant LNCaP:C42 prostate cancer cells and the effect on gene expression and splicing determined.

ORGANISM(S): Homo sapiens

PROVIDER: GSE210130 | GEO | 2023/01/04

REPOSITORIES: GEO

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The METTL3 RNA methyltransferase regulates androgen signalling in castrate resistant prostate cancer cells

Project description:iRNAs targeting METTL3 used to deplete METTL3 in castrate resistant LNCaP:C42 and 22Rv1 prosate cancer cells and the effect on basal and androgen regulated gene expression and splicing determined.

2022-11-02 | GSE194420 | GEO

A Cleavage Product METTL3a-(238-580) Mediates m6A Methyltransferase Complex Assembly and mTOR Activation

Project description:N6-methyladenosine (m6A) methylation of mRNA by the methyltransferase complex (MTC), with core components including METTL3-METTL14 heterodimers and Wilms’ tumor 1-associated protein (WTAP), contributes to breast tumorigenesis, but the mechanism of MTC assembly remains elusive. Here, we identify a novel cleaved form METTL3a (residues 239-580 of METTL3), that is highly expressed in breast cancer. Furthermore, we find that both METTL3a and full-length METTL3 are required for MTC assembly, RNA m6A deposition, as well as cancer cell proliferation. Mechanistically, we find that METTL3a is required for METTL3-METTL3 interaction, which is a prerequisite step for recruitment of WTAP in MTC assembly. Analysis of m6A sequencing data shows that depletion of METTL3a globally disrupts m6A methylation, and METTL3a mediates mTOR activation via m6A-mediated suppression of TMEM127 expression. Consequently, we find that METTL3 cleavage is mediated by proteasome in an mTOR-dependent manner, revealing positive regulatory feedback between METTL3a and mTOR signaling. Our findings reveal METTL3a as an important component for MTC assembly, and suggest the METTL3a-mTOR axis as a potential therapeutic target for breast cancer.

2023-08-16 | GSE213727 | GEO

m6A-seq profiling in Mettl3 Ctrl and dTAG13 mESCs [m6Aseq]

Project description:dTAG-13 treated METTL3_FKBP12(F36V) is sufficient to functionally deplete m6A methylation, that catalysed by METTL3/14 complex.

2020-09-15 | GSE154707 | GEO

The m6A methyltransferase Mettl3 deficiency attenuates hepatic stellate cell activation and liver fibrosis [m6A-RIP-seq]

Project description:m6A-RIP sequencing of primary hepatic stellate cells (HSCs) isolated from Control and HSC-specific Mettl3-knockout (Mettl3 cKO) mouse liver tissues.

2022-07-13 | GSE207908 | GEO

Proteomic analysis of castrate resistant prostate cancer cell lines

Project description:In this study the development of androgen independence in a cell model of disease was selected as a mirror of to the events at play in the development of Castrate Resistant Prostate Cancer in-vivo. LNCaP cells which are androgen dependent and androgen independent sublines; LNCaP-Abl and LNCaP-Abl-Hof were subject to extensive fractionation by 1-D SDS PAGE and accurate mass-high resolution mass spectrometry (Q Exactive) to identify proteins whose expression was changes significantly in response to androgen independent growth.

2022-02-28 | PXD000751 | Pride

BET inhibition abrogates global transcription via repressing m6A writer complex

Project description:To characterize the role of m6A modification in mediating BRD4-dependent biological functions, we established METTL3 reconstituting cells and treated with JQ1 BRD4 directs gene transcription through diverse mechanisms. Here, we specifically focused on BETi-dependent transcriptome or chestrated by m6A-mediated regulation. To this end, we first need to identify bona fide m6A-dependent transcripts via rescue experiments in METTL3-knockdown cells by adding back wild-type METTL3 or METTL3-CD mutant (a catalytically inactive form of METTL3)

2023-12-31 | GSE219191 | GEO

Redox Regulation of m6A Methyltransferase METTL3 in Human Beta Cells Controls the Innate Immune Response in Type 1 Diabetes

Project description:Label-free proteomics of S-nitrosylation on recombinant human METTL3 induced by S-Nitroso-N-acetyl-DL-penicillamine (SNAP) to determine redox-sensitive Cys sites

2023-03-24 | MSV000091547 | MassIVE

Identification and Characterization of the Mammalian Nuclear RNA N6-Adenosine Methyltransferase Core Complex

Project description:N6-methyladenosine (m6A) is the most prevalent internal modification found in mammalian messenger and non-coding RNAs. The discoveries of functionally significant demethylases that reverse this methylation as well as the recently revealed m6A distributions in mammalian transcriptomes strongly indicate regulatory functions of this modification. Here we report the identification and characterization of the mammalian nuclear RNA N6-adenosine methyltransferase core (RNMTC) complex. Besides METTL3, a methyltransferase which was the only known component of RNMTC in the past, we discovered that a previously uncharacterized methyltransferase, METTL14, exhibits a N6-adenosine methyltransferase activity higher than METTL3. Together with WTAP, the third component that dramatically affects the cellular m6A level, these three proteins form the core complex that orchestrates m6A deposition on mammalian nuclear RNA. Biochemistry assays, imaging experiments, as well as transcriptome-wide analyses of the binding sites and their effects on m6A methylation support methylation function and reveal new insights of RNMTC. PAR-CLIP and m6A-seq in HeLa cells

2013-11-29 | E-GEOD-46705 | biostudies-arrayexpress

m6A methyltransferase METTL3 regulates Tfh cell differentiation

Project description:m6A methyltransferase METTL3 regulates Tfh cell differentiation

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Determine the methyltransferase activities of METTL3, METTL14, and METTL16 via MeRIP-seq (m6A-seq)

Project description:All the three enzymes, METTL3, METTL14, and METTL16, belong to methyltransferase like (METTL) family and possess the ability to deposit N6-methyladenosine (m6A) in mRNA. Via immunofluorescence staining and wertern blotting, we discovered either METTL3 or METTL14 mainly localize in nuclear, but METTL16 localizes in both cytosol and nuclei. To compare the m6A methyltransferase activities of the three m6A 'writers' and better understand their differences, MeRIP-seq (m6A-seq) was conducted with poly(A) RNAs isolaed from HEK293T cells with METTL3, 14 and 16 knockout.

2021-10-28 | GSE182607 | GEO

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