A Method to Monitor the Introduction of Posttranscriptional Modifications in tRNAs with NMR Spectroscopy.
In extract NMR
NMR spectroscopy
Posttranscriptional modifications
RNA modifications
RNA transcription
Time-resolved NMR
Transfer RNA
Yeast
Yeast extract
tRNA
Journal
Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969
Informations de publication
Date de publication:
2021
2021
Historique:
entrez:
4
6
2021
pubmed:
5
6
2021
medline:
23
6
2021
Statut:
ppublish
Résumé
During their biosynthesis, transfer RNAs (tRNAs) are decorated with a large number of posttranscriptional chemical modifications. Methods to directly detect the introduction of posttranscriptional modifications during tRNA maturation are rare and do not provide information on the temporality of modification events. Here, we report a methodology, using NMR as a tool to monitor tRNA maturation in a nondisruptive and continuous fashion in cellular extracts. This method requires the production of substrate tRNA transcripts devoid of modifications and active cell extracts containing the necessary cellular enzymatic activities to modify RNA. The present protocol describes these different aspects of our method and reports the time-resolved NMR monitoring of the yeast tRNA
Identifiants
pubmed: 34085253
doi: 10.1007/978-1-0716-1374-0_19
doi:
Substances chimiques
RNA
63231-63-0
RNA, Transfer
9014-25-9
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
307-323Références
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