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ExoMeg1: a new exonuclease from metagenomic library

DNA repair mechanisms are responsible for maintaining the integrity of DNA and are essential to life. However, our knowledge of DNA repair mechanisms is based on model organisms such as Escherichia coli, and little is known about free living and uncultured microorganisms. In this study, a functional...

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Autores principales: Silva-Portela, Rita C. B., Carvalho, Fabíola M., Pereira, Carolina P. M., de Souza-Pinto, Nadja C., Modesti, Mauro, Fuchs, Robert P., Agnez-Lima, Lucymara F.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4750427/
https://www.ncbi.nlm.nih.gov/pubmed/26815639
http://dx.doi.org/10.1038/srep19712
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author Silva-Portela, Rita C. B.
Carvalho, Fabíola M.
Pereira, Carolina P. M.
de Souza-Pinto, Nadja C.
Modesti, Mauro
Fuchs, Robert P.
Agnez-Lima, Lucymara F.
author_facet Silva-Portela, Rita C. B.
Carvalho, Fabíola M.
Pereira, Carolina P. M.
de Souza-Pinto, Nadja C.
Modesti, Mauro
Fuchs, Robert P.
Agnez-Lima, Lucymara F.
author_sort Silva-Portela, Rita C. B.
collection PubMed
description DNA repair mechanisms are responsible for maintaining the integrity of DNA and are essential to life. However, our knowledge of DNA repair mechanisms is based on model organisms such as Escherichia coli, and little is known about free living and uncultured microorganisms. In this study, a functional screening was applied in a metagenomic library with the goal of discovering new genes involved in the maintenance of genomic integrity. One clone was identified and the sequence analysis showed an open reading frame homolog to a hypothetical protein annotated as a member of the Exo_Endo_Phos superfamily. This novel enzyme shows 3′-5′ exonuclease activity on single and double strand DNA substrates and it is divalent metal-dependent, EDTA-sensitive and salt resistant. The clone carrying the hypothetical ORF was able to complement strains deficient in recombination or base excision repair, suggesting that the new enzyme may be acting on the repair of single strand breaks with 3′ blockers, which are substrates for these repair pathways. Because this is the first report of an enzyme obtained from a metagenomic approach showing exonuclease activity, it was named ExoMeg1. The metagenomic approach has proved to be a useful tool for identifying new genes of uncultured microorganisms.
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spelling pubmed-47504272016-02-18 ExoMeg1: a new exonuclease from metagenomic library Silva-Portela, Rita C. B. Carvalho, Fabíola M. Pereira, Carolina P. M. de Souza-Pinto, Nadja C. Modesti, Mauro Fuchs, Robert P. Agnez-Lima, Lucymara F. Sci Rep Article DNA repair mechanisms are responsible for maintaining the integrity of DNA and are essential to life. However, our knowledge of DNA repair mechanisms is based on model organisms such as Escherichia coli, and little is known about free living and uncultured microorganisms. In this study, a functional screening was applied in a metagenomic library with the goal of discovering new genes involved in the maintenance of genomic integrity. One clone was identified and the sequence analysis showed an open reading frame homolog to a hypothetical protein annotated as a member of the Exo_Endo_Phos superfamily. This novel enzyme shows 3′-5′ exonuclease activity on single and double strand DNA substrates and it is divalent metal-dependent, EDTA-sensitive and salt resistant. The clone carrying the hypothetical ORF was able to complement strains deficient in recombination or base excision repair, suggesting that the new enzyme may be acting on the repair of single strand breaks with 3′ blockers, which are substrates for these repair pathways. Because this is the first report of an enzyme obtained from a metagenomic approach showing exonuclease activity, it was named ExoMeg1. The metagenomic approach has proved to be a useful tool for identifying new genes of uncultured microorganisms. Nature Publishing Group 2016-01-27 /pmc/articles/PMC4750427/ /pubmed/26815639 http://dx.doi.org/10.1038/srep19712 Text en Copyright © 2016, Macmillan Publishers Limited http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Silva-Portela, Rita C. B.
Carvalho, Fabíola M.
Pereira, Carolina P. M.
de Souza-Pinto, Nadja C.
Modesti, Mauro
Fuchs, Robert P.
Agnez-Lima, Lucymara F.
ExoMeg1: a new exonuclease from metagenomic library
title ExoMeg1: a new exonuclease from metagenomic library
title_full ExoMeg1: a new exonuclease from metagenomic library
title_fullStr ExoMeg1: a new exonuclease from metagenomic library
title_full_unstemmed ExoMeg1: a new exonuclease from metagenomic library
title_short ExoMeg1: a new exonuclease from metagenomic library
title_sort exomeg1: a new exonuclease from metagenomic library
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4750427/
https://www.ncbi.nlm.nih.gov/pubmed/26815639
http://dx.doi.org/10.1038/srep19712
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