m6Am-seq reveals the dynamic m6Am methylation in the human transcriptome

Hanxiao Sun, Kai Li, Xiaoting Zhang, Jun’e Liu, Meiling Zhang, Haowei Meng and Chengqi Yi ()
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Hanxiao Sun: Peking University
Kai Li: Peking University
Xiaoting Zhang: Peking University
Jun’e Liu: Peking University
Meiling Zhang: Peking University
Haowei Meng: Peking University
Chengqi Yi: Peking University

Nature Communications, 2021, vol. 12, issue 1, 1-12

Abstract: Abstract N6,2′-O-dimethyladenosine (m6Am), a terminal modification adjacent to the mRNA cap, is a newly discovered reversible RNA modification. Yet, a specific and sensitive tool to directly map transcriptome-wide m6Am is lacking. Here, we report m6Am-seq, based on selective in vitro demethylation and RNA immunoprecipitation. m6Am-seq directly distinguishes m6Am and 5′-UTR N6-methyladenosine (m6A) and enables the identification of m6Am at single-base resolution and 5′-UTR m6A in the human transcriptome. Using m6Am-seq, we also find that m6Am and 5′-UTR m6A respond dynamically to stimuli, and identify key functional methylation sites that may facilitate cellular stress response. Collectively, m6Am-seq reveals the high-confidence m6Am and 5′-UTR m6A methylome and provides a robust tool for functional studies of the two epitranscriptomic marks.

Date: 2021
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DOI: 10.1038/s41467-021-25105-5

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