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The dnd operon for DNA phosphorothioation modification system in Escherichia coli is located in diverse genomic islands

BACKGROUND: Strains of Escherichia coli that are non-typeable by pulsed-field gel electrophoresis (PFGE) due to in-gel degradation can influence their molecular epidemiological data. The DNA degradation phenotype (Dnd(+)) is mediated by the dnd operon that encode enzymes catalyzing the phosphorothio...

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Autores principales: Ho, Wing Sze, Ou, Hong-Yu, Yeo, Chew Chieng, Thong, Kwai Lin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4373003/
https://www.ncbi.nlm.nih.gov/pubmed/25879448
http://dx.doi.org/10.1186/s12864-015-1421-8
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author Ho, Wing Sze
Ou, Hong-Yu
Yeo, Chew Chieng
Thong, Kwai Lin
author_facet Ho, Wing Sze
Ou, Hong-Yu
Yeo, Chew Chieng
Thong, Kwai Lin
author_sort Ho, Wing Sze
collection PubMed
description BACKGROUND: Strains of Escherichia coli that are non-typeable by pulsed-field gel electrophoresis (PFGE) due to in-gel degradation can influence their molecular epidemiological data. The DNA degradation phenotype (Dnd(+)) is mediated by the dnd operon that encode enzymes catalyzing the phosphorothioation of DNA, rendering the modified DNA susceptible to oxidative cleavage during a PFGE run. In this study, a PCR assay was developed to detect the presence of the dnd operon in Dnd(+)E. coli strains and to improve their typeability. Investigations into the genetic environments of the dnd operon in various E. coli strains led to the discovery that the dnd operon is harboured in various diverse genomic islands. RESULTS: The dndBCDE genes (dnd operon) were detected in all Dnd(+)E. coli strains by PCR. The addition of thiourea improved the typeability of Dnd(+)E. coli strains to 100% using PFGE and the Dnd(+) phenotype can be observed in both clonal and genetically diverse E. coli strains. Genomic analysis of 101 dnd operons from genome sequences of Enterobacteriaceae revealed that the dnd operons of the same bacterial species were generally clustered together in the phylogenetic tree. Further analysis of dnd operons of 52 E. coli genomes together with their respective immediate genetic environments revealed a total of 7 types of genetic organizations, all of which were found to be associated with genomic islands designated dnd-encoding GIs. The dnd-encoding GIs displayed mosaic structure and the genomic context of the 7 islands (with 1 representative genome from each type of genetic organization) were also highly variable, suggesting multiple recombination events. This is also the first report where two dnd operons were found within a strain although the biological implication is unknown. Surprisingly, dnd operons were frequently found in pathogenic E. coli although their link with virulence has not been explored. CONCLUSION: Genomic islands likely play an important role in facilitating the horizontal gene transfer of the dnd operons in E. coli with 7 different types of islands discovered so far. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12864-015-1421-8) contains supplementary material, which is available to authorized users.
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spelling pubmed-43730032015-03-26 The dnd operon for DNA phosphorothioation modification system in Escherichia coli is located in diverse genomic islands Ho, Wing Sze Ou, Hong-Yu Yeo, Chew Chieng Thong, Kwai Lin BMC Genomics Research Article BACKGROUND: Strains of Escherichia coli that are non-typeable by pulsed-field gel electrophoresis (PFGE) due to in-gel degradation can influence their molecular epidemiological data. The DNA degradation phenotype (Dnd(+)) is mediated by the dnd operon that encode enzymes catalyzing the phosphorothioation of DNA, rendering the modified DNA susceptible to oxidative cleavage during a PFGE run. In this study, a PCR assay was developed to detect the presence of the dnd operon in Dnd(+)E. coli strains and to improve their typeability. Investigations into the genetic environments of the dnd operon in various E. coli strains led to the discovery that the dnd operon is harboured in various diverse genomic islands. RESULTS: The dndBCDE genes (dnd operon) were detected in all Dnd(+)E. coli strains by PCR. The addition of thiourea improved the typeability of Dnd(+)E. coli strains to 100% using PFGE and the Dnd(+) phenotype can be observed in both clonal and genetically diverse E. coli strains. Genomic analysis of 101 dnd operons from genome sequences of Enterobacteriaceae revealed that the dnd operons of the same bacterial species were generally clustered together in the phylogenetic tree. Further analysis of dnd operons of 52 E. coli genomes together with their respective immediate genetic environments revealed a total of 7 types of genetic organizations, all of which were found to be associated with genomic islands designated dnd-encoding GIs. The dnd-encoding GIs displayed mosaic structure and the genomic context of the 7 islands (with 1 representative genome from each type of genetic organization) were also highly variable, suggesting multiple recombination events. This is also the first report where two dnd operons were found within a strain although the biological implication is unknown. Surprisingly, dnd operons were frequently found in pathogenic E. coli although their link with virulence has not been explored. CONCLUSION: Genomic islands likely play an important role in facilitating the horizontal gene transfer of the dnd operons in E. coli with 7 different types of islands discovered so far. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12864-015-1421-8) contains supplementary material, which is available to authorized users. BioMed Central 2015-03-17 /pmc/articles/PMC4373003/ /pubmed/25879448 http://dx.doi.org/10.1186/s12864-015-1421-8 Text en © Ho et al.; licensee BioMed Central. 2015 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research Article
Ho, Wing Sze
Ou, Hong-Yu
Yeo, Chew Chieng
Thong, Kwai Lin
The dnd operon for DNA phosphorothioation modification system in Escherichia coli is located in diverse genomic islands
title The dnd operon for DNA phosphorothioation modification system in Escherichia coli is located in diverse genomic islands
title_full The dnd operon for DNA phosphorothioation modification system in Escherichia coli is located in diverse genomic islands
title_fullStr The dnd operon for DNA phosphorothioation modification system in Escherichia coli is located in diverse genomic islands
title_full_unstemmed The dnd operon for DNA phosphorothioation modification system in Escherichia coli is located in diverse genomic islands
title_short The dnd operon for DNA phosphorothioation modification system in Escherichia coli is located in diverse genomic islands
title_sort dnd operon for dna phosphorothioation modification system in escherichia coli is located in diverse genomic islands
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4373003/
https://www.ncbi.nlm.nih.gov/pubmed/25879448
http://dx.doi.org/10.1186/s12864-015-1421-8
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