Time-resolved NMR monitoring of tRNA maturation.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 31358763.
- Also identified by DOI 10.1038/s41467-019-11356-w and PMC identifier 6662845.
- Licence recorded as CC BY.
- The licence permits redistribution, so the abstract is shown in full and the full text is available from the publisher.
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 tRNA<sup>Phe</sup> 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 m<sup>1</sup>A58 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.
Medical subject headings
- Magnetic Resonance Spectroscopy
- RNA Processing, Post-Transcriptional
- RNA, Transfer
- Saccharomyces cerevisiae Proteins
- tRNA Methyltransferases