Time-resolved NMR monitoring of tRNA maturation

Pierre Barraud (), Alexandre Gato, Matthias Heiss, Marjorie Catala, Stefanie Kellner and Carine Tisné ()
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Pierre Barraud: Institut de biologie physico-chimique
Alexandre Gato: Institut de biologie physico-chimique
Matthias Heiss: Ludwig Maximilians University Munich
Marjorie Catala: Institut de biologie physico-chimique
Stefanie Kellner: Ludwig Maximilians University Munich
Carine Tisné: Institut de biologie physico-chimique

Nature Communications, 2019, vol. 10, issue 1, 1-14

Abstract: Abstract Although the biological importance of post-transcriptional RNA modifications in gene expression is widely appreciated, methods to directly detect their introduction during RNA biosynthesis are rare and do not easily provide information on the temporal nature of events. Here, we introduce the application of NMR spectroscopy to observe the maturation of tRNAs in cell extracts. By following the maturation of yeast tRNAPhe with time-resolved NMR measurements, we show that modifications are introduced in a defined sequential order, and that the chronology is controlled by cross-talk between modification events. In particular, we show that a strong hierarchy controls the introduction of the T54, Ψ55 and m1A58 modifications in the T-arm, and we demonstrate that the modification circuits identified in yeast extract with NMR also impact the tRNA modification process in living cells. The NMR-based methodology presented here could be adapted to investigate different aspects of tRNA maturation and RNA modifications in general.

Date: 2019
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DOI: 10.1038/s41467-019-11356-w

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