Characterization of CRISPR-Cas systems in the Ralstonia solanacearum species complex

Detalhes bibliográficos
Autor(a) principal: Xavier, André da Silva
Data de Publicação: 2019
Outros Autores: Almeida, Juliana Cristina Fraleon de, Melo, Alessandra Gonçalves de, Rousseau, Geneviève M., Tremblay, Denise M., Rezende, Rafael Reis de, Moineau, Sylvain, Alfenas‐Zerbini, Poliane
Tipo de documento: Artigo
Idioma: eng
Título da fonte: LOCUS Repositório Institucional da UFV
Texto Completo: https://doi.org/10.1111/mpp.12750
http://www.locus.ufv.br/handle/123456789/23952
Resumo: Clustered regularly interspaced short palindromic repeats (CRISPRs) are composed of an array of short DNA repeat sequences separated by unique spacer sequences that are flanked by associated (Cas) genes. CRISPR-Cas systems are found in the genomes of several microbes and can act as an adaptive immune mechanism against invading foreign nucleic acids, such as phage genomes. Here, we studied the CRISPRCas systems in plant-pathogenic bacteria of the Ralstonia sola- nacearum species complex (RSSC). A CRISPR-Cas system was found in 31% of RSSC genomes present in public databases. Specifically, CRISPR-Cas types I-E and II-C were found, with I-E being the most common. The presence of the same CRISPRCas types in distinct Ralstonia phylotypes and species suggests the acquisition of the system by a common ancestor before Ralstonia species segregation. In addition, a Cas1 phylogeny (I-E type) showed a perfect geographical segregation of phylotypes, supporting an ancient acquisition. Ralstonia solanacearum strains CFBP2957 and K60 T were challenged with a virulent phage, and the CRISPR arrays of bacteriophage insensitive mutants (BIMs) were analysed. No new spacer acquisition was detected in the analysed BIMs. The functionality of the CRISPR-Cas interference step was also tested in R. solanacearum CFBP2957 using a spacer-protospacer adjacent motif (PAM) delivery system, and no resistance was observed against phage phiAP1. Our results show that the CRISPR-Cas system in R. solanacearum CFBP2957 is not its primary antiviral strategy.
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spelling Xavier, André da SilvaAlmeida, Juliana Cristina Fraleon deMelo, Alessandra Gonçalves deRousseau, Geneviève M.Tremblay, Denise M.Rezende, Rafael Reis deMoineau, SylvainAlfenas‐Zerbini, Poliane2019-03-14T18:06:29Z2019-03-14T18:06:29Z2019-0213643703https://doi.org/10.1111/mpp.12750http://www.locus.ufv.br/handle/123456789/23952Clustered regularly interspaced short palindromic repeats (CRISPRs) are composed of an array of short DNA repeat sequences separated by unique spacer sequences that are flanked by associated (Cas) genes. CRISPR-Cas systems are found in the genomes of several microbes and can act as an adaptive immune mechanism against invading foreign nucleic acids, such as phage genomes. Here, we studied the CRISPRCas systems in plant-pathogenic bacteria of the Ralstonia sola- nacearum species complex (RSSC). A CRISPR-Cas system was found in 31% of RSSC genomes present in public databases. Specifically, CRISPR-Cas types I-E and II-C were found, with I-E being the most common. The presence of the same CRISPRCas types in distinct Ralstonia phylotypes and species suggests the acquisition of the system by a common ancestor before Ralstonia species segregation. In addition, a Cas1 phylogeny (I-E type) showed a perfect geographical segregation of phylotypes, supporting an ancient acquisition. Ralstonia solanacearum strains CFBP2957 and K60 T were challenged with a virulent phage, and the CRISPR arrays of bacteriophage insensitive mutants (BIMs) were analysed. No new spacer acquisition was detected in the analysed BIMs. The functionality of the CRISPR-Cas interference step was also tested in R. solanacearum CFBP2957 using a spacer-protospacer adjacent motif (PAM) delivery system, and no resistance was observed against phage phiAP1. Our results show that the CRISPR-Cas system in R. solanacearum CFBP2957 is not its primary antiviral strategy.engMolecular Plant PathologyVolume 20, Issue 02, Pages 223-239, February 2019Adaptive immunityTranscriptional controlBacteria-virus interactionCharacterization of CRISPR-Cas systems in the Ralstonia solanacearum species complexinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfinfo:eu-repo/semantics/openAccessreponame:LOCUS Repositório Institucional da UFVinstname:Universidade Federal de Viçosa (UFV)instacron:UFVORIGINALartigo.pdfartigo.pdftexto completoapplication/pdf1198542https://locus.ufv.br//bitstream/123456789/23952/1/artigo.pdf1c2113451a6a55a24b8a4fe2587670dcMD51LICENSElicense.txtlicense.txttext/plain; charset=utf-81748https://locus.ufv.br//bitstream/123456789/23952/2/license.txt8a4605be74aa9ea9d79846c1fba20a33MD52123456789/239522019-03-14 15:06:57.409oai:locus.ufv.br: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Repositório InstitucionalPUBhttps://www.locus.ufv.br/oai/requestfabiojreis@ufv.bropendoar:21452019-03-14T18:06:57LOCUS Repositório Institucional da UFV - Universidade Federal de Viçosa (UFV)false
dc.title.en.fl_str_mv Characterization of CRISPR-Cas systems in the Ralstonia solanacearum species complex
title Characterization of CRISPR-Cas systems in the Ralstonia solanacearum species complex
spellingShingle Characterization of CRISPR-Cas systems in the Ralstonia solanacearum species complex
Xavier, André da Silva
Adaptive immunity
Transcriptional control
Bacteria-virus interaction
title_short Characterization of CRISPR-Cas systems in the Ralstonia solanacearum species complex
title_full Characterization of CRISPR-Cas systems in the Ralstonia solanacearum species complex
title_fullStr Characterization of CRISPR-Cas systems in the Ralstonia solanacearum species complex
title_full_unstemmed Characterization of CRISPR-Cas systems in the Ralstonia solanacearum species complex
title_sort Characterization of CRISPR-Cas systems in the Ralstonia solanacearum species complex
author Xavier, André da Silva
author_facet Xavier, André da Silva
Almeida, Juliana Cristina Fraleon de
Melo, Alessandra Gonçalves de
Rousseau, Geneviève M.
Tremblay, Denise M.
Rezende, Rafael Reis de
Moineau, Sylvain
Alfenas‐Zerbini, Poliane
author_role author
author2 Almeida, Juliana Cristina Fraleon de
Melo, Alessandra Gonçalves de
Rousseau, Geneviève M.
Tremblay, Denise M.
Rezende, Rafael Reis de
Moineau, Sylvain
Alfenas‐Zerbini, Poliane
author2_role author
author
author
author
author
author
author
dc.contributor.author.fl_str_mv Xavier, André da Silva
Almeida, Juliana Cristina Fraleon de
Melo, Alessandra Gonçalves de
Rousseau, Geneviève M.
Tremblay, Denise M.
Rezende, Rafael Reis de
Moineau, Sylvain
Alfenas‐Zerbini, Poliane
dc.subject.pt-BR.fl_str_mv Adaptive immunity
Transcriptional control
Bacteria-virus interaction
topic Adaptive immunity
Transcriptional control
Bacteria-virus interaction
description Clustered regularly interspaced short palindromic repeats (CRISPRs) are composed of an array of short DNA repeat sequences separated by unique spacer sequences that are flanked by associated (Cas) genes. CRISPR-Cas systems are found in the genomes of several microbes and can act as an adaptive immune mechanism against invading foreign nucleic acids, such as phage genomes. Here, we studied the CRISPRCas systems in plant-pathogenic bacteria of the Ralstonia sola- nacearum species complex (RSSC). A CRISPR-Cas system was found in 31% of RSSC genomes present in public databases. Specifically, CRISPR-Cas types I-E and II-C were found, with I-E being the most common. The presence of the same CRISPRCas types in distinct Ralstonia phylotypes and species suggests the acquisition of the system by a common ancestor before Ralstonia species segregation. In addition, a Cas1 phylogeny (I-E type) showed a perfect geographical segregation of phylotypes, supporting an ancient acquisition. Ralstonia solanacearum strains CFBP2957 and K60 T were challenged with a virulent phage, and the CRISPR arrays of bacteriophage insensitive mutants (BIMs) were analysed. No new spacer acquisition was detected in the analysed BIMs. The functionality of the CRISPR-Cas interference step was also tested in R. solanacearum CFBP2957 using a spacer-protospacer adjacent motif (PAM) delivery system, and no resistance was observed against phage phiAP1. Our results show that the CRISPR-Cas system in R. solanacearum CFBP2957 is not its primary antiviral strategy.
publishDate 2019
dc.date.accessioned.fl_str_mv 2019-03-14T18:06:29Z
dc.date.available.fl_str_mv 2019-03-14T18:06:29Z
dc.date.issued.fl_str_mv 2019-02
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv https://doi.org/10.1111/mpp.12750
http://www.locus.ufv.br/handle/123456789/23952
dc.identifier.issn.none.fl_str_mv 13643703
identifier_str_mv 13643703
url https://doi.org/10.1111/mpp.12750
http://www.locus.ufv.br/handle/123456789/23952
dc.language.iso.fl_str_mv eng
language eng
dc.relation.ispartofseries.pt-BR.fl_str_mv Volume 20, Issue 02, Pages 223-239, February 2019
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Molecular Plant Pathology
publisher.none.fl_str_mv Molecular Plant Pathology
dc.source.none.fl_str_mv reponame:LOCUS Repositório Institucional da UFV
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instname_str Universidade Federal de Viçosa (UFV)
instacron_str UFV
institution UFV
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