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Identification of a 2′-O-Methyluridine Nucleoside Hydrolase Using the Metagenomic Libraries

Ribose methylation is among the most ubiquitous modifications found in RNA. 2′-O-methyluridine is found in rRNA, snRNA, snoRNA and tRNA of Archaea, Bacteria, and Eukaryota. Moreover, 2′-O-methylribonucleosides are promising starting materials for the production of nucleic acid-based drugs. Despite t...

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Autores principales: Aučynaitė, Agota, Rutkienė, Rasa, Tauraitė, Daiva, Meškys, Rolandas, Urbonavičius, Jaunius
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6278475/
https://www.ncbi.nlm.nih.gov/pubmed/30405065
http://dx.doi.org/10.3390/molecules23112904
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author Aučynaitė, Agota
Rutkienė, Rasa
Tauraitė, Daiva
Meškys, Rolandas
Urbonavičius, Jaunius
author_facet Aučynaitė, Agota
Rutkienė, Rasa
Tauraitė, Daiva
Meškys, Rolandas
Urbonavičius, Jaunius
author_sort Aučynaitė, Agota
collection PubMed
description Ribose methylation is among the most ubiquitous modifications found in RNA. 2′-O-methyluridine is found in rRNA, snRNA, snoRNA and tRNA of Archaea, Bacteria, and Eukaryota. Moreover, 2′-O-methylribonucleosides are promising starting materials for the production of nucleic acid-based drugs. Despite the countless possibilities of practical use for the metabolic enzymes associated with methylated nucleosides, there are very few reports regarding the metabolic fate and enzymes involved in the metabolism of 2′-O-alkyl nucleosides. The presented work focuses on the cellular degradation of 2′-O-methyluridine. A novel enzyme was found using a screening strategy that employs Escherichia coli uracil auxotroph and the metagenomic libraries. A 2′-O-methyluridine hydrolase (RK9NH) has been identified together with an aldolase (RK9DPA)—forming a part of a probable gene cluster that is involved in the degradation of 2′-O-methylated nucleosides. The RK9NH is functional in E. coli uracil auxotroph and in vitro. The RK9NH nucleoside hydrolase could be engineered to enzymatically produce 2′-O-methylated nucleosides that are of great demand as raw materials for production of nucleic acid-based drugs. Moreover, RK9NH nucleoside hydrolase converts 5-fluorouridine, 5-fluoro-2′-deoxyuridine and 5-fluoro-2′-O-methyluridine into 5-fluorouracil, which suggests it could be employed in cancer therapy.
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spelling pubmed-62784752018-12-13 Identification of a 2′-O-Methyluridine Nucleoside Hydrolase Using the Metagenomic Libraries Aučynaitė, Agota Rutkienė, Rasa Tauraitė, Daiva Meškys, Rolandas Urbonavičius, Jaunius Molecules Article Ribose methylation is among the most ubiquitous modifications found in RNA. 2′-O-methyluridine is found in rRNA, snRNA, snoRNA and tRNA of Archaea, Bacteria, and Eukaryota. Moreover, 2′-O-methylribonucleosides are promising starting materials for the production of nucleic acid-based drugs. Despite the countless possibilities of practical use for the metabolic enzymes associated with methylated nucleosides, there are very few reports regarding the metabolic fate and enzymes involved in the metabolism of 2′-O-alkyl nucleosides. The presented work focuses on the cellular degradation of 2′-O-methyluridine. A novel enzyme was found using a screening strategy that employs Escherichia coli uracil auxotroph and the metagenomic libraries. A 2′-O-methyluridine hydrolase (RK9NH) has been identified together with an aldolase (RK9DPA)—forming a part of a probable gene cluster that is involved in the degradation of 2′-O-methylated nucleosides. The RK9NH is functional in E. coli uracil auxotroph and in vitro. The RK9NH nucleoside hydrolase could be engineered to enzymatically produce 2′-O-methylated nucleosides that are of great demand as raw materials for production of nucleic acid-based drugs. Moreover, RK9NH nucleoside hydrolase converts 5-fluorouridine, 5-fluoro-2′-deoxyuridine and 5-fluoro-2′-O-methyluridine into 5-fluorouracil, which suggests it could be employed in cancer therapy. MDPI 2018-11-07 /pmc/articles/PMC6278475/ /pubmed/30405065 http://dx.doi.org/10.3390/molecules23112904 Text en © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Aučynaitė, Agota
Rutkienė, Rasa
Tauraitė, Daiva
Meškys, Rolandas
Urbonavičius, Jaunius
Identification of a 2′-O-Methyluridine Nucleoside Hydrolase Using the Metagenomic Libraries
title Identification of a 2′-O-Methyluridine Nucleoside Hydrolase Using the Metagenomic Libraries
title_full Identification of a 2′-O-Methyluridine Nucleoside Hydrolase Using the Metagenomic Libraries
title_fullStr Identification of a 2′-O-Methyluridine Nucleoside Hydrolase Using the Metagenomic Libraries
title_full_unstemmed Identification of a 2′-O-Methyluridine Nucleoside Hydrolase Using the Metagenomic Libraries
title_short Identification of a 2′-O-Methyluridine Nucleoside Hydrolase Using the Metagenomic Libraries
title_sort identification of a 2′-o-methyluridine nucleoside hydrolase using the metagenomic libraries
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6278475/
https://www.ncbi.nlm.nih.gov/pubmed/30405065
http://dx.doi.org/10.3390/molecules23112904
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