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Title:
Transcriptome-wide mapping reveals reversible and dynamic N1-methyladenosine methylome | Nature Chemical Biology
Description:
A method called m1A-ID-seq, which involves antibody enrichment and reverse transcriptase–based sequencing of N1-methyladenosine (m1A) RNA modifications, reveals that m1A is a reversible mRNA modification that is abundant in the 5′ UTRs of human genes. N1-Methyladenosine (m1A) is a prevalent post-transcriptional RNA modification, yet little is known about its abundance, topology and dynamics in mRNA. Here, we show that m1A is prevalent in Homo sapiens mRNA, which shows an m1A/A ratio of ∼0.02%. We develop the m1A-ID-seq technique, based on m1A immunoprecipitation and the inherent ability of m1A to stall reverse transcription, as a means for transcriptome-wide m1A profiling. m1A-ID-seq identifies 901 m1A peaks (from 600 genes) in mRNA and noncoding RNA and reveals a prominent feature, enrichment in the 5′ untranslated region of mRNA transcripts, that is distinct from the pattern for N6-methyladenosine, the most abundant internal mammalian mRNA modification. Moreover, m1A in mRNA is reversible by ALKBH3, a known DNA/RNA demethylase. Lastly, we show that m1A methylation responds dynamically to stimuli, and we identify hundreds of stress-induced m1A sites. Collectively, our approaches allow comprehensive analysis of m1A modification and provide tools for functional studies of potential epigenetic regulation via the reversible and dynamic m1A methylation.
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N1-Methyladenosine (m1A) is a prevalent post-transcriptional RNA modification, yet little is known about its abundance, topology and dynamics in mRNA. Here, we show that m1A is prevalent in Homo sapiens mRNA, which shows an m1A/A ratio of â¼0.02%. We develop the m1A-ID-seq technique, based on m1A immunoprecipitation and the inherent ability of m1A to stall reverse transcription, as a means for transcriptome-wide m1A profiling. m1A-ID-seq identifies 901 m1A peaks (from 600 genes) in mRNA and noncoding RNA and reveals a prominent feature, enrichment in the 5â² untranslated region of mRNA transcripts, that is distinct from the pattern for N6-methyladenosine, the most abundant internal mammalian mRNA modification. Moreover, m1A in mRNA is reversible by ALKBH3, a known DNA/RNA demethylase. Lastly, we show that m1A methylation responds dynamically to stimuli, and we identify hundreds of stress-induced m1A sites. Collectively, our approaches allow comprehensive analysis of m1A modification and provide tools for functional studies of potential epigenetic regulation via the reversible and dynamic m1A methylation.
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name:Peking-Tsinghua Center for Life Sciences, Peking University, Beijing, China
name:State Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, Beijing, China
name:Peking-Tsinghua Center for Life Sciences, Peking University, Beijing, China
name:State Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, Beijing, China
name:Peking-Tsinghua Center for Life Sciences, Peking University, Beijing, China
name:Academy for Advanced Interdisciplinary Studies, Peking University, Beijing, China
name:State Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, Beijing, China
name:Peking-Tsinghua Center for Life Sciences, Peking University, Beijing, China
name:Academy for Advanced Interdisciplinary Studies, Peking University, Beijing, China
name:State Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, Beijing, China
name:Peking-Tsinghua Center for Life Sciences, Peking University, Beijing, China
name:Department of Chemical Biology, Synthetic and Functional Biomolecules Center, College of Chemistry and Molecular Engineering, Peking University, Beijing, China
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