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Two novel proteins with opposing m6A writing functions interact with HAKAI [meRIP-seq]

ABSTRACT: Two novel proteins with opposing m6A writing functions interact with HAKAI [meRIP-seq]

PROVIDER: PRJNA730562 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR14570256_1.fastq.gz	Fastqsanger.gz
	SRR14570256_2.fastq.gz	Fastqsanger.gz
	SRR14570257_1.fastq.gz	Fastqsanger.gz
	SRR14570257_2.fastq.gz	Fastqsanger.gz
	SRR14570258_1.fastq.gz	Fastqsanger.gz

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Two novel proteins with opposing m6A writing functions interact with HAKAI [RNA-seq]

Project description:Two novel proteins with opposing m6A writing functions interact with HAKAI [RNA-seq]

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Two novel proteins with opposing m6A writing functions interact with HAKAI [meRIP-seq]

Project description:Using in vivo interaction proteomics, two HAKAI-interacting zinc finger proteins, HIZ1 and HIZ2, were discovered as novel components of the Arabidopsis m6A writer complex. HAKAI is required for the interaction between HIZ1 and MTA. Whilst HIZ1 knockout plants have normal levels of m6A, plants in which it is overexpressed show reduced methylation. In addition, HIZ1 was found to be involved in root hair development upon auxin transport inhibition in a HAKAI-dependent manner. Mutant plants lacking HIZ2 are viable but have an 85% reduction in m6A abundance and show severe developmental defects. Our findings suggest that HIZ1 appears to be a HAKAI-dependent negative regulator of m6A deposition and HIZ2 is a novel and essential member of the Arabidopsis m6A writer complex.

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Two novel proteins with opposing m6A writing functions interact with HAKAI [RNA-seq]

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Two zinc finger proteins with functions in m6A writing interact with HAKAI

Project description:Two zinc finger proteins with functions in m6A writing interact with HAKAI

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Two zinc finger proteins with functions in m6A writing interact with HAKAI

Project description:The methyltransferase complex (m6A writer), which catalyzes the deposition of N6-methyladenosine (m6A) in mRNAs, is highly conserved across most eukaryotic organisms, but its components and interactions between them are still far from fully understood. Using in vivo interaction proteomics, two HAKAI-interacting zinc finger proteins, HIZ1 and HIZ2, were discovered as novel components of the Arabidopsis m6A writer complex. HAKAI is required for the interaction between HIZ1 and MTA. Whilst HIZ1 knockout plants have normal levels of m6A, plants in which it is overexpressed show reduced methylation. In addition, HIZ1 was found to be involved in root hair development upon auxin transport inhibition in a HAKAI-dependent manner. Mutant plants lacking HIZ2 are viable but have an 85% reduction in m6A abundance and show severe developmental defects. Our findings suggest that HIZ1 appears to be a HAKAI-dependent negative regulator of m6A deposition and HIZ2 is a novel and essential member of the Arabidopsis m6A writer complex.

2022-02-09 | PXD026287 | Pride

m6A-RIP-seq of mRNA in yw, Mettl3, Mettl14 and Hakai flies

Project description:In order to obtain a stringent m6A methylome in Drosophila adults, we performed m6A-RIP-seq in yw control (male and female), Mettl3 (male), Mettl14 (male) and Hakai (male) mutant flies. We find that the effective m6A modification, which depends on the writer complex, is mostly distributed in 5’ UTR and near start codon in Drosophila, in contrast to the mammalian system. We define a set of high-confident m6A methylation sites shared by Mettl3, Mettl14 and Hakai, indicating that Hakai is a core component of the m6A writer complex. We also find differential methylation pattern in certain loci between male and female flies.

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Project description:The mRNA m6A reader YTHDF2 is overexpressed in a broad spectrum of human acute myeloid leukemias (AML). To understand the role of YTHDF2 in AML, we generated m6A meRIP-seq libraries form Ythdf2fl/fl (Ythdf2CTL) pre-leukemic cells.

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m6A meRIP-Seq from Ythdf2CKO pre-leukemic cells

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Hakai is required for stabilizing core components of the m6A mRNA methylation machinery

Project description:N6-methyladenosine RNA (m6A) is the most abundant internal modification on mRNA which influences most steps of mRNA metabolism and is involved in several biological functions, including circadian clock, metabolism and embryonic stem cell differentiation. The E3 ubiquitin ligase Hakai was previously found in complex with components of the m6A methylation machinery in plants and mammalian cells but its precise function remained to be investigated. Here we show that Hakai is a conserved component of the methyltransferase complex in Drosophila. Its depletion results in reduced m6A levels and altered m6A-dependent functions including sex determination. We show that its ubiquitination domain is required for dimerisation and interaction with other members of the m6A machinery, while its catalytic activity seems dispensable. Finally, we demonstrate that the loss of Hakai destabilizes the level of several subunits of the methyltransferase complex, resulting in impaired m6A deposition. Thus, our work adds new functional and molecular insights into the mechanism of the m6A mRNA writer complex.

2021-04-26 | GSE158663 | GEO

m6A-meRIP sequencing

Project description:m6A-meRIP sequencing

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