[PMID]: | 28703578 |
[Au] Autor: | Wulff TF; Argüello RJ; Molina Jordàn M; Roura Frigolé H; Hauquier G; Filonava L; Camacho N; Gatti E; Pierre P; Ribas de Pouplana L; Torres AG |
[Ad] Endereço: | Institute for Research in Biomedicine (IRB Barcelona), The Barcelona Institute of Science and Technology , Parc Científic de Barcelona, C/Baldiri Reixac 10, 08028 Barcelona, Catalonia, Spain. |
[Ti] Título: | Detection of a Subset of Posttranscriptional Transfer RNA Modifications in Vivo with a Restriction Fragment Length Polymorphism-Based Method. |
[So] Source: | Biochemistry;56(31):4029-4038, 2017 Aug 08. |
[Is] ISSN: | 1520-4995 |
[Cp] País de publicação: | United States |
[La] Idioma: | eng |
[Ab] Resumo: | 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. |
[Mh] Termos MeSH primário: |
Adenosina Desaminase/metabolismo Modelos Biológicos Polimorfismo de Fragmento de Restrição Processamento Pós-Transcricional do RNA RNA de Transferência de Alanina/metabolismo RNA de Transferência de Treonina/metabolismo
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[Mh] Termos MeSH secundário: |
Adenosina/metabolismo Adenosina Desaminase/química Adenosina Desaminase/genética Análise do Polimorfismo de Comprimento de Fragmentos Amplificados Pareamento de Bases Biologia Computacional Desaminação Sistemas Especialistas Células HeLa Seres Humanos Concentração de Íons de Hidrogênio Inosina/metabolismo Interferência de RNA RNA Interferente Pequeno/metabolismo RNA de Transferência de Alanina/antagonistas & inibidores RNA de Transferência de Treonina/antagonistas & inibidores RNA de Transferência de Valina/antagonistas & inibidores RNA de Transferência de Valina/metabolismo Transcrição Reversa Especificidade por Substrato
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[Pt] Tipo de publicação: | JOURNAL ARTICLE; VALIDATION STUDIES |
[Nm] Nome de substância:
| 0 (RNA, Small Interfering); 0 (RNA, Transfer, Ala); 0 (RNA, Transfer, Thr); 0 (RNA, Transfer, Val); 5A614L51CT (Inosine); EC 3.5.4.4 (ADAT2 protein, human); EC 3.5.4.4 (Adenosine Deaminase); K72T3FS567 (Adenosine) |
[Em] Mês de entrada: | 1708 |
[Cu] Atualização por classe: | 170818 |
[Lr] Data última revisão:
| 170818 |
[Sb] Subgrupo de revista: | IM |
[Da] Data de entrada para processamento: | 170714 |
[St] Status: | MEDLINE |
[do] DOI: | 10.1021/acs.biochem.7b00324 |
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