2026-04-17T05:38:43Zhttps://recercat.cat/oai/request

oai:recercat.cat:2445/1149172025-12-04T19:32:37Zcom_2072_1057col_2072_478908col_2072_478917

00925njm 22002777a 4500 dc Wulff, Thomas F. author Argüello, Rafael J. author Molina Jordan, Marc author Roura Frigolé, Helena author Hauquier, Glenn author Filonava, Liudmila author Camacho, Noelia author Gatti, Evelina author Pierre, Philippe author Ribas de Pouplana, Lluís author Torres, Adrian G. author 2017-09-04T10:38:24Z 2018-07-13T22:01:22Z 2017-07-13 2017-08-07T11:55:05Z Transfer RNAs (tRNAs) are among the most heavily modified RNA species. Posttranscriptional tRNA modifications (ptRMs) play fundamental roles in modulating tRNA structure and function and are being increasingly linked to human physiology and disease. Detection of ptRMs is often challenging, expensive, and laborious. Restriction fragment length polymorphism (RFLP) analyses study the patterns of DNA cleavage after restriction enzyme treatment and have been used for the qualitative detection of modified bases on mRNAs. It is known that some ptRMs induce specific and reproducible base “mutations” when tRNAs are reverse transcribed. For example, inosine, which derives from the deamination of adenosine, is detected as a guanosine when an inosine-containing tRNA is reverse transcribed, amplified via polymerase chain reaction (PCR), and sequenced. ptRM-dependent base changes on reverse transcription PCR amplicons generated as a consequence of the reverse transcription reaction might create or abolish endonuclease restriction sites. The suitability of RFLP for the detection and/or quantification of ptRMs has not been studied thus far. Here we show that different ptRMs can be detected at specific sites of different tRNA types by RFLP. For the examples studied, we show that this approach can reliably estimate the modification status of the sample, a feature that can be useful in the study of the regulatory role of tRNA modifications in gene expression. ADN Expressió gènica DNA Gene expression Detection of a Subset of Posttranscriptional Transfer RNA Modifications in Vivo with a Restriction Fragment Length Polymorphism-Based Method