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Genomewide m6A mapping uncovers dynamic changes in the m6A epitranscriptome of cisplatin-treated apoptotic HeLa cells

ABSTRACT: Genomewide m6A mapping uncovers dynamic changes in the m6A epitranscriptome of cisplatin-treated apoptotic HeLa cells

PROVIDER: PRJNA779444 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR16912861.fastq.gz	Fastqsanger.gz
	SRR16912862.fastq.gz	Fastqsanger.gz
	SRR16912863.fastq.gz	Fastqsanger.gz
	SRR16912864.fastq.gz	Fastqsanger.gz
	SRR16912865.fastq.gz	Fastqsanger.gz

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Genomewide m6A mapping uncovers dynamic changes in the m6A epitranscriptome of cisplatin-treated apoptotic HeLa cells

Project description:We have performed a epitranscriptomics study in which we first tretaed DMSO as a negative control and cisplatin in HeLa cells. Total RNA was isolated from control as well as CP treated cells and apoptotic rate was determined by flow cytometry. Total RNAs were subjected to miCLIP to identify differentially m6A methylated RNAs. We then transcriptionally analyzed apoptotic genes which have differential m6A methylation in CP tretament by qPCR. Afterthat, candidate genes were examined at the level of translation by polysome fractioantion assay.

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Project description:N6-methyladenosine (m6A) is a widespread reversible chemical modification of RNAs, implicated in many aspects of RNA metabolism. Little quantitative information exists as to either how many transcript copies of particular genes are m6A modified (âm6A levelsâ), or the relationship of m6A modification(s) to alternative RNA isoforms. To deconvolute the m6A epitranscriptome, we developed m6A level and isoform-characterization sequencing (m6A-LAIC-seq). We found that cells exhibit a broad range of non-stoichiometric m6A levels with cell type specificity. At the level of isoform characterization, we discovered widespread differences in use of tandem alternative polyadenylation (APA) sites by methylated and nonmethylated transcript isoforms of individual genes. Strikingly, there is a strong bias for methylated transcripts to be coupled with proximal APA sites, resulting in shortened 3â untranslated regions (3â-UTRs), while nonmethylated transcript isoforms tend to use distal APA sites. m6A-LAIC-seq yields a new perspective on transcriptome complexity and links APA usage to m6A modifications. m6A-LAIC-seq of H1-ESC and GM12878 cell lines, each cell line has two replicates

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