Time-resolved NMR monitoring of tRNA maturation

Autor: Stefanie Kellner, Matthias Heiss, Carine Tisné, Alexandre Gato, Marjorie Catala, Pierre Barraud
Přispěvatelé: Expression Génétique Microbienne (EGM (UMR_8261 / FRE_3630)), Institut de biologie physico-chimique (IBPC), Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS), Ludwig-Maximilians-Universität München (LMU), Institut de biologie physico-chimique (IBPC (FR_550))
Jazyk: angličtina
Rok vydání: 2019
Předmět:
0301 basic medicine
Models
Molecular

TRNA modification
Magnetic Resonance Spectroscopy
Saccharomyces cerevisiae Proteins
Time Factors
Science
General Physics and Astronomy
02 engineering and technology
Saccharomyces cerevisiae
010402 general chemistry
01 natural sciences
General Biochemistry
Genetics and Molecular Biology

Article
03 medical and health sciences
RNA
Transfer
Phe

RNA
Transfer

Gene expression
RNA Processing
Post-Transcriptional

lcsh:Science
ComputingMilieux_MISCELLANEOUS
030304 developmental biology
0303 health sciences
tRNA Methyltransferases
Multidisciplinary
Base Sequence
Chemistry
RNA
General Chemistry
Nuclear magnetic resonance spectroscopy
021001 nanoscience & nanotechnology
RNA modification
Yeast
0104 chemical sciences
[SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry
Molecular Biology/Biomolecules [q-bio.BM]

Nucleic acids
[SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry
Molecular Biology/Biomolecules [q-bio.BM]

030104 developmental biology
Order (biology)
Biochemistry
Transfer RNA
Nucleic Acid Conformation
lcsh:Q
0210 nano-technology
RNA biosynthesis
TRNA maturation
Solution-state NMR
Zdroj: Nature Communications
Nature Communications, Nature Publishing Group, 2019, 10 (1), ⟨10.1038/s41467-019-11356-w⟩
Nature Communications, Vol 10, Iss 1, Pp 1-14 (2019)
ISSN: 2041-1723
DOI: 10.1038/s41467-019-11356-w⟩
Popis: 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.
Transfer RNA (tRNA) is regulated by RNA modifications. Here the authors employ time-resolved NMR to monitor modifications of yeast tRNAPhe in cellular extracts, revealing a sequential order and cross-talk between modifications.
Databáze: OpenAIRE