TY - JOUR
T1 - The m1A landscape on cytosolic and mitochondrial mRNA at single-base resolution
AU - Safra, Modi
AU - Sas-Chen, Aldema
AU - Nir, Ronit
AU - Winkler, Roni
AU - Nachshon, Aharon
AU - Bar-Yaacov, Dan
AU - Erlacher, Matthias
AU - Rossmanith, Walter
AU - Stern-Ginossar, Noam
AU - Schwartz, Schraga
N1 - Funding Information: Acknowledgements This project was supported by the Israel Science Foundation (543165), the European Research Council under the European Union’s Horizon 2020 research and innovation programme (grant agreement number 714023), by the Abisch-Frenkel-Stiftung, by research grants from The Abramson Family Center for Young Scientists, the David and Fela Shapell Family Foundation INCPM Fund for Preclinical Studies, the Estate of David Turner, and the Berlin Family Foundation New Scientist Fund.
PY - 2017/11/9
Y1 - 2017/11/9
N2 - Modifications on mRNA offer the potential of regulating mRNA fate post-transcriptionally. Recent studies suggested the widespread presence of N 1 -methyladenosine (m 1 A), 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 m 1 A at single-nucleotide resolution. Within the cytosol, m 1 A 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 m 1 A site in the mitochondrial ND5 mRNA, catalysed by TRMT10C, with methylation levels that are highly tissue specific and tightly developmentally controlled. m 1 A leads to translational repression, probably through a mechanism involving ribosomal scanning or translation. Our findings suggest that m 1 A 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 m 1 A levels was adopted as a potential means of post-transcriptional regulation.
AB - Modifications on mRNA offer the potential of regulating mRNA fate post-transcriptionally. Recent studies suggested the widespread presence of N 1 -methyladenosine (m 1 A), 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 m 1 A at single-nucleotide resolution. Within the cytosol, m 1 A 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 m 1 A site in the mitochondrial ND5 mRNA, catalysed by TRMT10C, with methylation levels that are highly tissue specific and tightly developmentally controlled. m 1 A leads to translational repression, probably through a mechanism involving ribosomal scanning or translation. Our findings suggest that m 1 A 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 m 1 A levels was adopted as a potential means of post-transcriptional regulation.
KW - Adenosine/analogs & derivatives
KW - Base Pairing
KW - Cytosol/metabolism
KW - Electron Transport Complex I/biosynthesis
KW - Gene Expression Regulation
KW - HEK293 Cells
KW - Humans
KW - Membrane Proteins/genetics
KW - Methylation
KW - Methyltransferases/metabolism
KW - Mitochondria/genetics
KW - Mitochondrial Proteins/biosynthesis
KW - Organ Specificity
KW - Protein Biosynthesis
KW - RNA, Messenger/chemistry
KW - RNA, Mitochondrial
KW - RNA, Transfer/metabolism
KW - RNA/chemistry
KW - Transcriptome
KW - tRNA Methyltransferases/genetics
UR - http://www.scopus.com/inward/record.url?scp=85033486019&partnerID=8YFLogxK
U2 - https://doi.org/10.1038/nature24456
DO - https://doi.org/10.1038/nature24456
M3 - مقالة
C2 - 29072297
SN - 0028-0836
VL - 551
SP - 251
EP - 255
JO - Nature
JF - Nature
IS - 7679
ER -