Evidence for the Active Role of a Novel Nuclease from Helicobacter pylori in the Horizontal Transfer of Genetic Information

Autores
O'Rourke, Eyleen J.; Pinto, A. Viviana; Petroni, E. Alejandro; Tolmasky, Marcelo E.; Ielpi, Luis
Año de publicación
2004
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Helicobacter pylori is a gram-negative bacterium that colonizes the human stomach, causes gastritis, and is associated with ulcers and gastric cancer. H. pylori is naturally competent for transformation. Natural genetic transformation is believed to be essential for the genetic plasticity observed in this species. While the relevance of horizontal gene transfer in H. pylori adaptiveness and antibiotic resistance is well documented, the DNA transformation machinery components are barely known. No enzymatic activity associated with the transformation process has been determined experimentally and described. We isolated, microsequenced, and cloned a major DNA nuclease from H. pylori. This protein, encoded by the open reading frame hp0323, was expressed in Escherichia coli. The purified protein, NucT, has a cation-independent thermostable nuclease activity that preferentially cleaves single-stranded DNA. NucT is associated with the membrane. NucT-deficient H. pylori strains are one or more orders of magnitude less efficient than the parental strain for transformation with either chromosomal or self-replicating plasmid DNA. To the best of our knowledge, NucT is the first nuclease identified in a gram-negative natural transformation system, and its existence suggests that there is a mechanism of DNA processing and uptake similar to the mechanisms in well-studied gram-positive systems.
Fil: O'Rourke, Eyleen J.. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina. Instituto Pasteur; Francia. Centre National de la Recherche Scientifique; Francia
Fil: Pinto, A. Viviana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina. Centre National de la Recherche Scientifique; Francia
Fil: Petroni, E. Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina
Fil: Tolmasky, Marcelo E.. California State University; Estados Unidos
Fil: Ielpi, Luis. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina
Materia
Helicobacter pylori
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/43014
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/43014
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Evidence for the Active Role of a Novel Nuclease from Helicobacter pylori in the Horizontal Transfer of Genetic InformationO'Rourke, Eyleen J.Pinto, A. VivianaPetroni, E. AlejandroTolmasky, Marcelo E.Ielpi, LuisHelicobacter pylorihttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Helicobacter pylori is a gram-negative bacterium that colonizes the human stomach, causes gastritis, and is associated with ulcers and gastric cancer. H. pylori is naturally competent for transformation. Natural genetic transformation is believed to be essential for the genetic plasticity observed in this species. While the relevance of horizontal gene transfer in H. pylori adaptiveness and antibiotic resistance is well documented, the DNA transformation machinery components are barely known. No enzymatic activity associated with the transformation process has been determined experimentally and described. We isolated, microsequenced, and cloned a major DNA nuclease from H. pylori. This protein, encoded by the open reading frame hp0323, was expressed in Escherichia coli. The purified protein, NucT, has a cation-independent thermostable nuclease activity that preferentially cleaves single-stranded DNA. NucT is associated with the membrane. NucT-deficient H. pylori strains are one or more orders of magnitude less efficient than the parental strain for transformation with either chromosomal or self-replicating plasmid DNA. To the best of our knowledge, NucT is the first nuclease identified in a gram-negative natural transformation system, and its existence suggests that there is a mechanism of DNA processing and uptake similar to the mechanisms in well-studied gram-positive systems.Fil: O'Rourke, Eyleen J.. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina. Instituto Pasteur; Francia. Centre National de la Recherche Scientifique; FranciaFil: Pinto, A. Viviana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina. Centre National de la Recherche Scientifique; FranciaFil: Petroni, E. Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; ArgentinaFil: Tolmasky, Marcelo E.. California State University; Estados UnidosFil: Ielpi, Luis. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; ArgentinaAmerican Society for Microbiology2004-04info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/43014O'Rourke, Eyleen J.; Pinto, A. Viviana; Petroni, E. Alejandro; Tolmasky, Marcelo E.; Ielpi, Luis; Evidence for the Active Role of a Novel Nuclease from Helicobacter pylori in the Horizontal Transfer of Genetic Information; American Society for Microbiology; Journal Of Bacteriology; 186; 9; 4-2004; 2586-25930021-91931098-5530CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/http://jb.asm.org/content/186/9/2586.longinfo:eu-repo/semantics/altIdentifier/doi/10.1128/JB.186.9.2586-2593.2004info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T10:05:15Zoai:ri.conicet.gov.ar:11336/43014instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-03 10:05:15.609CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Evidence for the Active Role of a Novel Nuclease from Helicobacter pylori in the Horizontal Transfer of Genetic Information
title Evidence for the Active Role of a Novel Nuclease from Helicobacter pylori in the Horizontal Transfer of Genetic Information
spellingShingle Evidence for the Active Role of a Novel Nuclease from Helicobacter pylori in the Horizontal Transfer of Genetic Information
O'Rourke, Eyleen J.
Helicobacter pylori
title_short Evidence for the Active Role of a Novel Nuclease from Helicobacter pylori in the Horizontal Transfer of Genetic Information
title_full Evidence for the Active Role of a Novel Nuclease from Helicobacter pylori in the Horizontal Transfer of Genetic Information
title_fullStr Evidence for the Active Role of a Novel Nuclease from Helicobacter pylori in the Horizontal Transfer of Genetic Information
title_full_unstemmed Evidence for the Active Role of a Novel Nuclease from Helicobacter pylori in the Horizontal Transfer of Genetic Information
title_sort Evidence for the Active Role of a Novel Nuclease from Helicobacter pylori in the Horizontal Transfer of Genetic Information
dc.creator.none.fl_str_mv O'Rourke, Eyleen J.
Pinto, A. Viviana
Petroni, E. Alejandro
Tolmasky, Marcelo E.
Ielpi, Luis
author O'Rourke, Eyleen J.
author_facet O'Rourke, Eyleen J.
Pinto, A. Viviana
Petroni, E. Alejandro
Tolmasky, Marcelo E.
Ielpi, Luis
author_role author
author2 Pinto, A. Viviana
Petroni, E. Alejandro
Tolmasky, Marcelo E.
Ielpi, Luis
author2_role author
author
author
author
dc.subject.none.fl_str_mv Helicobacter pylori
topic Helicobacter pylori
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Helicobacter pylori is a gram-negative bacterium that colonizes the human stomach, causes gastritis, and is associated with ulcers and gastric cancer. H. pylori is naturally competent for transformation. Natural genetic transformation is believed to be essential for the genetic plasticity observed in this species. While the relevance of horizontal gene transfer in H. pylori adaptiveness and antibiotic resistance is well documented, the DNA transformation machinery components are barely known. No enzymatic activity associated with the transformation process has been determined experimentally and described. We isolated, microsequenced, and cloned a major DNA nuclease from H. pylori. This protein, encoded by the open reading frame hp0323, was expressed in Escherichia coli. The purified protein, NucT, has a cation-independent thermostable nuclease activity that preferentially cleaves single-stranded DNA. NucT is associated with the membrane. NucT-deficient H. pylori strains are one or more orders of magnitude less efficient than the parental strain for transformation with either chromosomal or self-replicating plasmid DNA. To the best of our knowledge, NucT is the first nuclease identified in a gram-negative natural transformation system, and its existence suggests that there is a mechanism of DNA processing and uptake similar to the mechanisms in well-studied gram-positive systems.
Fil: O'Rourke, Eyleen J.. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina. Instituto Pasteur; Francia. Centre National de la Recherche Scientifique; Francia
Fil: Pinto, A. Viviana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina. Centre National de la Recherche Scientifique; Francia
Fil: Petroni, E. Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina
Fil: Tolmasky, Marcelo E.. California State University; Estados Unidos
Fil: Ielpi, Luis. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina
description Helicobacter pylori is a gram-negative bacterium that colonizes the human stomach, causes gastritis, and is associated with ulcers and gastric cancer. H. pylori is naturally competent for transformation. Natural genetic transformation is believed to be essential for the genetic plasticity observed in this species. While the relevance of horizontal gene transfer in H. pylori adaptiveness and antibiotic resistance is well documented, the DNA transformation machinery components are barely known. No enzymatic activity associated with the transformation process has been determined experimentally and described. We isolated, microsequenced, and cloned a major DNA nuclease from H. pylori. This protein, encoded by the open reading frame hp0323, was expressed in Escherichia coli. The purified protein, NucT, has a cation-independent thermostable nuclease activity that preferentially cleaves single-stranded DNA. NucT is associated with the membrane. NucT-deficient H. pylori strains are one or more orders of magnitude less efficient than the parental strain for transformation with either chromosomal or self-replicating plasmid DNA. To the best of our knowledge, NucT is the first nuclease identified in a gram-negative natural transformation system, and its existence suggests that there is a mechanism of DNA processing and uptake similar to the mechanisms in well-studied gram-positive systems.
publishDate 2004
dc.date.none.fl_str_mv 2004-04
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/43014
O'Rourke, Eyleen J.; Pinto, A. Viviana; Petroni, E. Alejandro; Tolmasky, Marcelo E.; Ielpi, Luis; Evidence for the Active Role of a Novel Nuclease from Helicobacter pylori in the Horizontal Transfer of Genetic Information; American Society for Microbiology; Journal Of Bacteriology; 186; 9; 4-2004; 2586-2593
0021-9193
1098-5530
CONICET Digital
CONICET
url http://hdl.handle.net/11336/43014
identifier_str_mv O'Rourke, Eyleen J.; Pinto, A. Viviana; Petroni, E. Alejandro; Tolmasky, Marcelo E.; Ielpi, Luis; Evidence for the Active Role of a Novel Nuclease from Helicobacter pylori in the Horizontal Transfer of Genetic Information; American Society for Microbiology; Journal Of Bacteriology; 186; 9; 4-2004; 2586-2593
0021-9193
1098-5530
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/http://jb.asm.org/content/186/9/2586.long
info:eu-repo/semantics/altIdentifier/doi/10.1128/JB.186.9.2586-2593.2004
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv American Society for Microbiology
publisher.none.fl_str_mv American Society for Microbiology
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.13397

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