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Intact DNA strands can be found in fossils, while scientists sometimes struggle to keep RNA intact under laboratory conditions. The structural variations between RNA and DNA underlie the differences in their stability and longevity. Because DNA is double-stranded, it is inherently more stable. The single-stranded structure of RNA is less stable but also more flexible and can form weak internal bonds. Additionally, most RNAs in the cell are relatively short, while DNA can be up to 250 million...
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A Method for Measuring RNA N6-methyladenosine Modifications in Cells and Tissues
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N6-methyladenosine modification destabilizes developmental regulators in embryonic stem cells.

Yang Wang1, Yue Li2, Julia I Toth1

  • 1Tumor Initiation and Maintenance Program, NCI-designated Cancer Center, Sanford Burnham Medical Research Institute, La Jolla, California 92037, USA.

Nature Cell Biology
|January 8, 2014
PubMed
Summary

Methyltransferase-like 3 (Mettl3) and Mettl14 proteins synergistically control N(6)-methyladenosine (m(6)A) RNA methylation in mammals. This m(6)A modification is crucial for maintaining mouse embryonic stem cell self-renewal and ground state.

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Area of Science:

  • Molecular Biology
  • Epigenetics
  • Stem Cell Biology

Background:

  • N(6)-methyladenosine (m(6)A) is the most prevalent internal modification of eukaryotic messenger RNA.
  • While m(6)A's roles in yeast and plants are known, its function in mammals is largely unexplored.
  • m(6)A modification is observed in thousands of mammalian mRNAs and lncRNAs, and m(6)A demethylases are vital for mammalian homeostasis and fertility.

Purpose of the Study:

  • To identify the mammalian proteins responsible for m(6)A formation.
  • To investigate the function of m(6)A modification in mammalian cells, specifically in embryonic stem cells.
  • To elucidate the regulatory mechanisms involving m(6)A in maintaining stem cell identity.

Main Methods:

  • Identification and characterization of methyltransferase-like 3 (Mettl3) and Mettl14 proteins.
  • Knockdown of Mettl3 and Mettl14 in mouse embryonic stem cells (mESCs).
  • Analysis of m(6)A RNA methylation levels, mRNA stability, gene expression, and involvement of HuR and microRNA pathways.

Main Results:

  • Mettl3 and Mettl14 function synergistically as the primary m(6)A methyltransferases in mammalian cells.
  • Knockdown of Mettl3 or Mettl14 in mESCs resulted in a loss of m(6)A methylation and self-renewal capacity.
  • m(6)A methylation inversely correlates with mRNA stability and gene expression for numerous transcripts, including developmental regulators, mediated by HuR and microRNA pathways.

Conclusions:

  • Mettl3 and Mettl14 form a complex essential for m(6)A RNA methylation in mammalian cells.
  • m(6)A modification regulated by Mettl3/Mettl14 is critical for maintaining the ground state and self-renewal of mESCs.
  • This m(6)A-mediated gene regulatory mechanism is widespread, affecting thousands of transcripts across various cell types.