A Method to Monitor the Introduction of Posttranscriptional Modifications in tRNAs with NMR Spectroscopy.
Methods Mol Biol
; 2298: 307-323, 2021.
Article
en En
| MEDLINE
| ID: mdl-34085253
ABSTRACT
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 tRNAPhe maturation as an example. The NMR-based methodology presented here could be adapted to investigate diverse features in tRNA maturation.
Palabras clave
Texto completo:
1
Banco de datos:
MEDLINE
Asunto principal:
ARN de Transferencia
/
Espectroscopía de Resonancia Magnética
/
Procesamiento Postranscripcional del ARN
Idioma:
En
Revista:
Methods Mol Biol
Asunto de la revista:
BIOLOGIA MOLECULAR
Año:
2021
Tipo del documento:
Article
País de afiliación:
Francia