Transcriptome-wide maps of m6A RNA methylation marks in murine liver
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ABSTRACT:
PROVIDER: PRJNA744537 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR15064825.fastq.gz | Fastqsanger.gz | |||
| SRR15064826.fastq.gz | Fastqsanger.gz | |||
| SRR15064827.fastq.gz | Fastqsanger.gz | |||
| SRR15064828.fastq.gz | Fastqsanger.gz | |||
| SRR15064829.fastq.gz | Fastqsanger.gz |
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Project description:The intestinal microbiota modulates host physiology and gene expression via mechanisms that are not fully understood. A recently discovered layer of gene expression regulation is N6-methyladenosine (m6A) and N6,2′ -O-dimethyladenosine (m6Am) modifications of mRNA. To unveil if these epitranscriptomic marks are affected by the gut microbiota, we performed methylated RNA-immunoprecipitation and sequencing (MeRIP-seq) to examine m6A-modifications in transcripts of mice displaying either a conventional, or a modified, or no gut microbiota and discovered that the microbiota has a strong influence on m6A- modifications in the cecum, and also, albeit to a lesser extent, in the liver, affecting pathways related to metabolism, inflammatory and antimicrobial responses . We furthermore analysed expression levels of several known writer and eraser enzymes and found the methyltransferase Mettl16 to be downregulated in absence of a microbiota. As a consequence, one of its targets, the S-adenosyl methionine synthase Mat2a was less expressed in mice without gut flora. We furthermore show that distinct commensal bacteria, Akkermansia muciniphila, Lactobacillus plantarum can affect specific m6A modifications. Together, we report here epitranscriptomic modifications as an additional level of interaction in the complex interplay between commensal bacteria and their host.
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Project description:We report the application of sequencing technology for high-throughput profiling of mRNA m6A methylation in mice colonic epithelial cells with or without mettl14 deletion. By obtaining over thirty billion bases of sequence from m6A antibody immunoprecipitated mRNA, we generated m6A methylation maps of mouse colonic epithelial cells with or with mettl14 deletion. We find that GsdmC2, GsdmC3, GadmC3 mRNA methylation is robustly decreased in colonic epithelial cells upon mettl14 deletion, leading to the decrease of Gsdmc expression.
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Project description:We report the application of single-molecule-based sequencing technology for high-throughput profiling of histone modifications in mammalian cells. By obtaining over four billion bases of sequence from chromatin immunoprecipitated DNA, we generated genome-wide chromatin-state maps of mouse liver. We find that lysine 4 and lysine 27 trimethylation effectively discriminates genes that are expressed, poised for expression, or stably repressed, and therefore reflect cell state and lineage potential. Lysine 36 trimethylation marks primary coding and non-coding transcripts, facilitating gene annotation. Trimethylation of lysine 9 and lysine 20 is detected at satellite, telomeric and active long-terminal repeats, and can spread into proximal unique sequences. Lysine 4 and lysine 9 trimethylation marks imprinting control regions. Finally, we show that chromatin state can be read in an allele-specific manner by using single nucleotide polymorphisms. This study provides a framework for the application of comprehensive chromatin profiling towards characterization of diverse mammalian cell populations.
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Project description:Epigenetic modifications are known to profoundly affect the development and behavior of social insects. In the well-known caste differentiation process of honeybee (Apis mellifera), female larvae with identical genomes are fed royal jellydifferently and develop into either normal workers or into very large, long-lived, and extremely fecund queens, and the queen-worker asymmetry of honeybee is known to be result largely to differential genomic imprinting during larval development that involves DNA methylation-based regulation. The discovery of reversible N6-methyladenosine (m6A) RNA methylation modification has defined a new era for RNA-metabolism-related genetic regulation, yet much remains unknown about m6A-mediated post-transcriptional regulatory mechanisms. Here, we report the first honeybee RNA m6A methylome. Specifically, we used the m6A-seq technique to examine the RNA m6A methylomes of honeybee larvae, including queen and worker larvae at multiple instar stages. We identified multiple conserved features of m6A methylation machinery and transcriptome-wide m6A distribution trends among insect species, and observed that m6A marks exert functions in regulating caste differentiation, with apparently particularly strong functional impacts on fifth instar worker larvae. Functional annotation of differentially methylated candidate caste-differentiation-related transcripts revealed many known regulators of caste differentiation (e.g. ILP-2, p110, PI3K, and JHAMT etc.) as well as the widely-studied Vitellogenin gene, which has not previously been implicated in caste differentiation. As ever-more regulatory roles for m6A marks are discovered, honeybees may become an excellent model studying the biology of such epi-transcriptomic regulatory systems, from embryonic development through holometabolous caste-specific development and on towards behavior and the emergent social hierarchies underlying eusociality in animals.
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Project description:m6A is a ubiquitous RNA modification in eukaryotes. Transcriptome-wide m6A patterns in Arabidopsis have been assayed recently. However, m6A differential patterns among organs have not been well characterized. The goal of the study is to comprehensively analyze m6A patterns of numerous types of RNAs, the relationship between transcript level and m6A methylation extent, and m6A differential patterns among organs in Arabidopsis. In total, 18 libraries were sequneced. For the 3 organs: leaf, flower and root, each organ has mRNA-Seq, m6A-Seq and Input sequenced. And each sequence has 2 replicats.
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Project description:N6-methyl-adenosine (m6A) is the most abundant modification on messenger RNAs and is linked to human diseases, but its functions in mammalian development are poorly understood. Here we reveal the evolutionary conservation and function of m6A by mapping the m6A methylome in mouse and human embryonic stem cells. Thousands of messenger and long noncoding RNAs show conserved m6A modification, including transcripts encoding core pluripotency transcription factors. m6A is enriched over 3M-bM-^@M-^Y untranslated regions at defined sequence motifs, and marks unstable transcripts, including transcripts turned over upon differentiation. Genetic inactivation or depletion of mouse and human Mettl3, one of the m6A methylases, led to m6A erasure on select target genes, prolonged Nanog expression upon differentiation, and impaired ESCM-bM-^@M-^Ys exit from self-renewal towards differentiation into several lineages in vitro and in vivo. Thus, m6A is a mark of transcriptome flexibility required for stem cells to differentiate to specific lineages. Examing m6A modification differences in two different cell types
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