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A Method for Measuring RNA N6-methyladenosine Modifications in Cells and Tissues
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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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Widespread occurrence of N6-methyladenosine in bacterial mRNA

Xin Deng1, Kai Chen2, Guan-Zheng Luo2

  • 1TEDA Institute of Biological Sciences and Biotechnology, Nankai University, 23 Hongda Street, Tianjin 300457, P.R. China Key Laboratory of Molecular Microbiology and Technology, Ministry of Education, Tianjin 300071, P.R. China dengxindx@gmail.com.

Nucleic Acids Research
|June 13, 2015
PubMed

Insights

N(6)-methyladenosine (m(6)A) is a prevalent modification in bacterial messenger RNA (mRNA), distinct from eukaryotic patterns. This discovery opens new avenues for understanding bacterial gene regulation and RNA modification.

Area of Science:

  • Molecular Biology
  • RNA Biology
  • Microbiology

Background:

  • N(6)-methyladenosine (m(6)A) is the most abundant internal modification in eukaryotic messenger RNA (mRNA).
  • While m(6)A is found in bacterial ribosomal RNA, its presence in bacterial mRNA was previously elusive.
  • Recent advancements in identifying m(6)A regulators and methylomes in eukaryotes highlighted its regulatory roles.

Purpose of the Study:

  • To investigate the occurrence and abundance of m(6)A modification in bacterial mRNA.
  • To profile transcriptome-wide m(6)A patterns in key bacterial species.
  • To identify conserved m(6)A motifs and associated gene functions in bacteria.

Main Methods:

  • Utilized ultra-high pressure liquid chromatography coupled with triple-quadrupole tandem mass spectrometry (UHPLC-QQQ-MS/MS) to quantify the m(6)A/A ratio in bacterial mRNA.
  • Performed transcriptome-wide m(6)A profiling in *Escherichia coli* and *Pseudomonas aeruginosa*.
  • Analyzed m(6)A peak locations, consensus motifs, and performed functional enrichment analysis.

Main Results:

  • Demonstrated that m(6)A is an abundant mRNA modification across a wide range of bacterial species.
  • Revealed a conserved m(6)A pattern in *E. coli* and *P. aeruginosa* that differs from eukaryotic patterns.
  • Identified that most bacterial m(6)A peaks are located within open reading frames, featuring a conserved GCCAU motif.
  • Found that m(6)A-modified genes are predominantly involved in respiration, amino acid metabolism, stress response, and small RNA pathways.

Conclusions:

  • m(6)A is a widespread and abundant mRNA modification in bacteria, challenging previous assumptions.
  • The conserved m(6)A pattern and motif in bacteria suggest specific regulatory mechanisms.
  • Bacterial m(6)A likely plays significant roles in fundamental cellular processes such as metabolism and stress response.

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