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A Method for Measuring RNA N6-methyladenosine Modifications in Cells and Tissues
Published on: December 5, 2016
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Transcriptome-wide mapping reveals reversible and dynamic N(1)-methyladenosine methylome
Xiaoyu Li1,2, Xushen Xiong1,2,3, Kun Wang1,2
1State Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, Beijing, China.
Nature Chemical Biology
|February 11, 2016
Summary
N(1)-Methyladenosine (m(1)A) is a widespread RNA modification in human mRNA. This study introduces m(1)A-ID-seq to map m(1)A sites, revealing its dynamic and reversible nature.
Area of Science:
- Molecular Biology
- Epigenetics
- RNA Modifications
Background:
- N(1)-Methyladenosine (m(1)A) is a significant post-transcriptional RNA modification.
- Its abundance, distribution, and dynamics in mRNA remain largely unexplored.
Purpose of the Study:
- To investigate the prevalence and characteristics of m(1)A in Homo sapiens mRNA.
- To develop a method for transcriptome-wide m(1)A profiling.
- To explore the reversibility and dynamic regulation of m(1)A.
Main Methods:
- Development of m(1)A-ID-seq, a technique combining m(1)A immunoprecipitation and reverse transcription stalling.
- Transcriptome-wide profiling of m(1)A modifications.
- Assays to assess m(1)A reversibility by ALKBH3.
Main Results:
- m(1)A is prevalent in human mRNA with an m(1)A/A ratio of approximately 0.02%.
- m(1)A-ID-seq identified 901 m(1)A peaks in mRNA and noncoding RNA, with enrichment in the 5' untranslated region.
- m(1)A modification is reversible by ALKBH3 and dynamically responds to stimuli, with hundreds of stress-induced sites identified.
Conclusions:
- m(1)A is a dynamic and reversible RNA modification in human mRNA, distinct from N(6)-methyladenosine.
- The developed m(1)A-ID-seq technique enables comprehensive analysis of m(1)A.
- These findings provide tools for studying potential epigenetic regulation by m(1)A.
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