The dynamic epitranscriptome: N 6-methyladenosine and gene expression control. Gene expression regulation mediated through reversible m 6A RNA methylation. FMRP modulates neural differentiation through m 6A-dependent mRNA nuclear export. Recognition of RNA N 6-methyladenosine by IGF2BP proteins enhances mRNA stability and translation. Nuclear m 6A reader YTHDC1 regulates mRNA splicing. N 6-methyladenosine-dependent regulation of messenger RNA stability. ALKBH5 is a mammalian RNA demethylase that impacts RNA metabolism and mouse fertility. N 6-methyladenosine in nuclear RNA is a major substrate of the obesity-associated FTO. Mammalian WTAP is a regulatory subunit of the RNA N 6-methyladenosine methyltransferase. A METT元–METTL14 complex mediates mammalian nuclear RNA N 6-adenosine methylation. Purification and cDNA cloning of the AdoMet-binding subunit of the human mRNA ( N 6-adenosine)-methyltransferase. Post-transcriptional gene regulation by mRNA modifications. Dynamic RNA modifications in gene expression regulation. Epitranscriptome sequencing technologies: decoding RNA modifications. Detecting RNA modifications in the epitranscriptome: predict and validate. MODOMICS: a database of RNA modification pathways-2017 update. This method offers advantages in detecting clustered m 6A sites and holds promise to locate nuclear nascent RNA m 6A modifications. We identified a few thousand mRNA m 6A sites in human HeLa, HEK293T and mouse H2.35 cells, carried out a parallel comparison of m 6A-label-seq with available m 6A sequencing methods, and validated selected sites by an orthogonal method. We pinpointed the mRNA a 6A locations based on iodination-induced misincorporation at the opposite site in complementary DNA during reverse transcription. Cellular RNAs could therefore be metabolically modified with N 6-allyladenosine (a 6A) at supposed m 6A-generating adenosine sites. Human and mouse cells could be fed with a methionine analog, Se-allyl- l-selenohomocysteine, which substitutes the methyl group on the enzyme cofactor SAM with the allyl. Here, we report a metabolic labeling method to detect mRNA m 6A transcriptome-wide at base resolution, called ‘m 6A-label-seq’. Transcriptome-wide mapping of N 6-methyladenosine (m 6A) at base resolution remains an issue, impeding our understanding of m 6A roles at the nucleotide level.
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