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Published on: July 10, 2020
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Chemical Method to Sequence 5-Formylcytosine on RNA
Ang Li1, Xuemeng Sun1, A Emilia Arguello1
1Department of Chemistry, Princeton University, Princeton, New Jersey 08544, United States.
ACS Chemical Biology
|February 25, 2022
Summary
A new chemical sequencing method, Mal-Seq, detects 5-formylcytosine (f5C) RNA modifications. This technique reveals high levels of f5C in mammalian mitochondrial tRNAs, but its absence in lower eukaryotes, offering insights into epitranscriptomic evolution.
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- Epitranscriptomic RNA modifications regulate biological processes.
- Identifying RNA modifications at nucleotide resolution remains challenging.
Purpose of the Study:
- To develop a chemical sequencing method for detecting 5-formylcytosine (f5C) modifications in RNA.
- To investigate the prevalence of f5C in mitochondrial tRNA (mt-tRNA) across different species and tissues.
Main Methods:
- Developed Mal-Seq, a chemical labeling method using malononitrile to detect f5C residues.
- Mal-Seq converts f5C to C-to-T mutations for sequencing via reverse transcription and PCR.
- Applied Mal-Seq to analyze f5C at the wobble position of mt-tRNA(Met).
Main Results:
- Mal-Seq selectively and efficiently labels f5C modifications.
- High-level f5C modification was observed in mammalian mt-tRNA(Met).
- f5C modification was notably absent in lower eukaryotes' mt-tRNA(Met).
Conclusions:
- Mal-Seq provides a novel platform for characterizing the f5C epitranscriptome.
- The study illuminates the evolutionary landscape of mitochondrial tRNA modifications.
- Findings highlight significant differences in f5C prevalence between mammals and lower eukaryotes.
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