The m1A landscape on cytosolic and mitochondrial mRNA at single-base resolution

Nature. 2017 Nov 9;551(7679):251-255. doi: 10.1038/nature24456. Epub 2017 Oct 25.

Abstract

Modifications on mRNA offer the potential of regulating mRNA fate post-transcriptionally. Recent studies suggested the widespread presence of N1-methyladenosine (m1A), which disrupts Watson-Crick base pairing, at internal sites of mRNAs. These studies lacked the resolution of identifying individual modified bases, and did not identify specific sequence motifs undergoing the modification or an enzymatic machinery catalysing them, rendering it challenging to validate and functionally characterize putative sites. Here we develop an approach that allows the transcriptome-wide mapping of m1A at single-nucleotide resolution. Within the cytosol, m1A is present in a low number of mRNAs, typically at low stoichiometries, and almost invariably in tRNA T-loop-like structures, where it is introduced by the TRMT6/TRMT61A complex. We identify a single m1A site in the mitochondrial ND5 mRNA, catalysed by TRMT10C, with methylation levels that are highly tissue specific and tightly developmentally controlled. m1A leads to translational repression, probably through a mechanism involving ribosomal scanning or translation. Our findings suggest that m1A on mRNA, probably because of its disruptive impact on base pairing, leads to translational repression, and is generally avoided by cells, while revealing one case in mitochondria where tight spatiotemporal control over m1A levels was adopted as a potential means of post-transcriptional regulation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine / analogs & derivatives*
  • Adenosine / metabolism
  • Base Pairing
  • Cytosol / metabolism*
  • Electron Transport Complex I / biosynthesis
  • Electron Transport Complex I / genetics
  • Gene Expression Regulation
  • HEK293 Cells
  • Humans
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism
  • Methylation
  • Methyltransferases / metabolism
  • Mitochondria / genetics*
  • Mitochondrial Proteins / biosynthesis
  • Mitochondrial Proteins / genetics
  • Organ Specificity
  • Protein Biosynthesis
  • RNA / chemistry*
  • RNA / genetics
  • RNA / metabolism*
  • RNA, Messenger / chemistry*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism*
  • RNA, Mitochondrial
  • RNA, Transfer / metabolism
  • Transcriptome
  • tRNA Methyltransferases / genetics
  • tRNA Methyltransferases / metabolism

Substances

  • CRLS1 protein, human
  • Membrane Proteins
  • Mitochondrial Proteins
  • RNA, Messenger
  • RNA, Mitochondrial
  • mitochondrial messenger RNA
  • 1-methyladenosine
  • RNA
  • RNA, Transfer
  • ND5 protein, human
  • Methyltransferases
  • TRMT10c protein, human
  • tRNA Methyltransferases
  • TRM61 protein, human
  • Electron Transport Complex I
  • Adenosine