The CO2-dependence of Brucella ovis and Brucella abortus biovars is caused by defective carbonic anhydrases

Brucella bacteria cause brucellosis, a major zoonosis whose control requires efficient diagnosis and vaccines. Identification of classical Brucella spp. has traditionally relied on phenotypic characterization, including surface antigens and 5¿10% CO2 necessity for growth (CO2-dependence), a trait of...

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Autores: Pérez-Etayo, L. (Lara)|||/items/57b924a2-7204-4a4c-ad38-831daceb4453, Miguel, M.J. (María Jesús) de|||/items/4968c547-5897-4fb3-86c1-d860c9ba4861, Conde-Álvarez, R. (Raquel)|||/items/7a805559-3962-434b-9df9-ec243d61b5aa, Muñoz, P. (Pilar)|||/items/4e98f4c8-d3e4-4fb3-b781-07a0bcb652bc, Khames, M. (Mammar)|||/items/512703b8-e4a1-4355-8a75-5085e0ee9508, Iriarte-Cilveti, M. (Maite)|||/items/5b20b123-3967-4f07-b577-13d13aa57363, Moriyon, I. (Ignacio)|||/items/834b4ce9-a879-4ba7-8884-39d0ddd88c7b, Zuñiga-Ripa, A. (Amaia)|||/items/9aa6da2e-e507-4319-a12c-092436f70feb
Tipo de documento: artigo
Data de publicação:2018
País:España
Recursos:Universidad de Navarra
Repositório:Dadun. Depósito Académico Digital de la Universidad de Navarra
Idioma:inglês
OAI Identifier:oai:dadun.unav.edu:10171/53656
Acesso em linha:https://hdl.handle.net/10171/53656
Access Level:Acceso aberto
Palavra-chave:Brucella
Microbiología
Melitensis Rev-1 vaccine
Pyruvate phosphate dikinase
Outer-membrane proteins
GLPX fructose-1,6-bisphosphatases
Laboratory models
Escherichia-coli
Full virulence
Ambient air
Growth
Rams
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oai_identifier_str oai:dadun.unav.edu:10171/53656
network_acronym_str ES
network_name_str España
repository_id_str
spelling The CO2-dependence of Brucella ovis and Brucella abortus biovars is caused by defective carbonic anhydrasesPérez-Etayo, L. (Lara)|||/items/57b924a2-7204-4a4c-ad38-831daceb4453Miguel, M.J. (María Jesús) de|||/items/4968c547-5897-4fb3-86c1-d860c9ba4861Conde-Álvarez, R. (Raquel)|||/items/7a805559-3962-434b-9df9-ec243d61b5aaMuñoz, P. (Pilar)|||/items/4e98f4c8-d3e4-4fb3-b781-07a0bcb652bcKhames, M. (Mammar)|||/items/512703b8-e4a1-4355-8a75-5085e0ee9508Iriarte-Cilveti, M. (Maite)|||/items/5b20b123-3967-4f07-b577-13d13aa57363Moriyon, I. (Ignacio)|||/items/834b4ce9-a879-4ba7-8884-39d0ddd88c7bZuñiga-Ripa, A. (Amaia)|||/items/9aa6da2e-e507-4319-a12c-092436f70febBrucellaMicrobiologíaMelitensis Rev-1 vaccinePyruvate phosphate dikinaseOuter-membrane proteinsGLPX fructose-1,6-bisphosphatasesLaboratory modelsEscherichia-coliFull virulenceAmbient airGrowthRamsBrucella bacteria cause brucellosis, a major zoonosis whose control requires efficient diagnosis and vaccines. Identification of classical Brucella spp. has traditionally relied on phenotypic characterization, including surface antigens and 5¿10% CO2 necessity for growth (CO2-dependence), a trait of Brucella ovis and most Brucella abortus biovars 1¿4 strains. Although molecular tests are replacing phenotypic methods, CO2-dependence remains of interest as it conditions isolation and propagation and reflects Brucella metabolism, an area of active research. Here, we investigated the connection of CO2-dependence and carbonic anhydrases (CA), the enzymes catalyzing the hydration of CO2 to the bicarbonate used by anaplerotic and biosynthetic carboxylases. Based on the previous demonstration that B. suis carries two functional CAs (CAI and CAII), we analyzed the CA sequences of CO2-dependent and -independent brucellae and spontaneous mutants. The comparisons strongly suggested that CAII is not functional in CO2-dependent B. abortus and B. ovis, and that a modified CAII sequence explains the CO2-independent phenotype of spontaneous mutants. Then, by mutagenesis and heterologous plasmid complementation and chromosomal insertion we proved that CAI alone is enough to support CO2-independent growth of B. suis in rich media but not of B. abortus in rich media or B. suis in minimal media. Finally, we also found that insertion of a heterologous active CAII into B. ovis reverted the CO2-dependence but did not alter its virulence in the mouse model. These results allow a better understanding of central aspects of Brucella metabolism and, in the case of B. ovis, provide tools for large-scale production of diagnostic antigens and vaccines.Dadun. Depósito Académico Digital Universidad de Navarra20182018-09-0620182018-01-0120182018-01-01journal articlehttp://purl.org/coar/resource_type/c_6501info:eu-repo/semantics/articleapplication/pdfhttps://hdl.handle.net/10171/53656reponame:Dadun. Depósito Académico Digital de la Universidad de Navarrainstname:Universidad de NavarraInglésengopen accesshttp://purl.org/coar/access_right/c_abf2info:eu-repo/semantics/openAccessoai:dadun.unav.edu:10171/536562026-06-21T12:47:57Z
dc.title.none.fl_str_mv The CO2-dependence of Brucella ovis and Brucella abortus biovars is caused by defective carbonic anhydrases
title The CO2-dependence of Brucella ovis and Brucella abortus biovars is caused by defective carbonic anhydrases
spellingShingle The CO2-dependence of Brucella ovis and Brucella abortus biovars is caused by defective carbonic anhydrases
Pérez-Etayo, L. (Lara)|||/items/57b924a2-7204-4a4c-ad38-831daceb4453
Brucella
Microbiología
Melitensis Rev-1 vaccine
Pyruvate phosphate dikinase
Outer-membrane proteins
GLPX fructose-1,6-bisphosphatases
Laboratory models
Escherichia-coli
Full virulence
Ambient air
Growth
Rams
title_short The CO2-dependence of Brucella ovis and Brucella abortus biovars is caused by defective carbonic anhydrases
title_full The CO2-dependence of Brucella ovis and Brucella abortus biovars is caused by defective carbonic anhydrases
title_fullStr The CO2-dependence of Brucella ovis and Brucella abortus biovars is caused by defective carbonic anhydrases
title_full_unstemmed The CO2-dependence of Brucella ovis and Brucella abortus biovars is caused by defective carbonic anhydrases
title_sort The CO2-dependence of Brucella ovis and Brucella abortus biovars is caused by defective carbonic anhydrases
dc.creator.none.fl_str_mv Pérez-Etayo, L. (Lara)|||/items/57b924a2-7204-4a4c-ad38-831daceb4453
Miguel, M.J. (María Jesús) de|||/items/4968c547-5897-4fb3-86c1-d860c9ba4861
Conde-Álvarez, R. (Raquel)|||/items/7a805559-3962-434b-9df9-ec243d61b5aa
Muñoz, P. (Pilar)|||/items/4e98f4c8-d3e4-4fb3-b781-07a0bcb652bc
Khames, M. (Mammar)|||/items/512703b8-e4a1-4355-8a75-5085e0ee9508
Iriarte-Cilveti, M. (Maite)|||/items/5b20b123-3967-4f07-b577-13d13aa57363
Moriyon, I. (Ignacio)|||/items/834b4ce9-a879-4ba7-8884-39d0ddd88c7b
Zuñiga-Ripa, A. (Amaia)|||/items/9aa6da2e-e507-4319-a12c-092436f70feb
author Pérez-Etayo, L. (Lara)|||/items/57b924a2-7204-4a4c-ad38-831daceb4453
author_facet Pérez-Etayo, L. (Lara)|||/items/57b924a2-7204-4a4c-ad38-831daceb4453
Miguel, M.J. (María Jesús) de|||/items/4968c547-5897-4fb3-86c1-d860c9ba4861
Conde-Álvarez, R. (Raquel)|||/items/7a805559-3962-434b-9df9-ec243d61b5aa
Muñoz, P. (Pilar)|||/items/4e98f4c8-d3e4-4fb3-b781-07a0bcb652bc
Khames, M. (Mammar)|||/items/512703b8-e4a1-4355-8a75-5085e0ee9508
Iriarte-Cilveti, M. (Maite)|||/items/5b20b123-3967-4f07-b577-13d13aa57363
Moriyon, I. (Ignacio)|||/items/834b4ce9-a879-4ba7-8884-39d0ddd88c7b
Zuñiga-Ripa, A. (Amaia)|||/items/9aa6da2e-e507-4319-a12c-092436f70feb
author_role author
author2 Miguel, M.J. (María Jesús) de|||/items/4968c547-5897-4fb3-86c1-d860c9ba4861
Conde-Álvarez, R. (Raquel)|||/items/7a805559-3962-434b-9df9-ec243d61b5aa
Muñoz, P. (Pilar)|||/items/4e98f4c8-d3e4-4fb3-b781-07a0bcb652bc
Khames, M. (Mammar)|||/items/512703b8-e4a1-4355-8a75-5085e0ee9508
Iriarte-Cilveti, M. (Maite)|||/items/5b20b123-3967-4f07-b577-13d13aa57363
Moriyon, I. (Ignacio)|||/items/834b4ce9-a879-4ba7-8884-39d0ddd88c7b
Zuñiga-Ripa, A. (Amaia)|||/items/9aa6da2e-e507-4319-a12c-092436f70feb
author2_role author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Dadun. Depósito Académico Digital Universidad de Navarra
dc.subject.none.fl_str_mv Brucella
Microbiología
Melitensis Rev-1 vaccine
Pyruvate phosphate dikinase
Outer-membrane proteins
GLPX fructose-1,6-bisphosphatases
Laboratory models
Escherichia-coli
Full virulence
Ambient air
Growth
Rams
topic Brucella
Microbiología
Melitensis Rev-1 vaccine
Pyruvate phosphate dikinase
Outer-membrane proteins
GLPX fructose-1,6-bisphosphatases
Laboratory models
Escherichia-coli
Full virulence
Ambient air
Growth
Rams
description Brucella bacteria cause brucellosis, a major zoonosis whose control requires efficient diagnosis and vaccines. Identification of classical Brucella spp. has traditionally relied on phenotypic characterization, including surface antigens and 5¿10% CO2 necessity for growth (CO2-dependence), a trait of Brucella ovis and most Brucella abortus biovars 1¿4 strains. Although molecular tests are replacing phenotypic methods, CO2-dependence remains of interest as it conditions isolation and propagation and reflects Brucella metabolism, an area of active research. Here, we investigated the connection of CO2-dependence and carbonic anhydrases (CA), the enzymes catalyzing the hydration of CO2 to the bicarbonate used by anaplerotic and biosynthetic carboxylases. Based on the previous demonstration that B. suis carries two functional CAs (CAI and CAII), we analyzed the CA sequences of CO2-dependent and -independent brucellae and spontaneous mutants. The comparisons strongly suggested that CAII is not functional in CO2-dependent B. abortus and B. ovis, and that a modified CAII sequence explains the CO2-independent phenotype of spontaneous mutants. Then, by mutagenesis and heterologous plasmid complementation and chromosomal insertion we proved that CAI alone is enough to support CO2-independent growth of B. suis in rich media but not of B. abortus in rich media or B. suis in minimal media. Finally, we also found that insertion of a heterologous active CAII into B. ovis reverted the CO2-dependence but did not alter its virulence in the mouse model. These results allow a better understanding of central aspects of Brucella metabolism and, in the case of B. ovis, provide tools for large-scale production of diagnostic antigens and vaccines.
publishDate 2018
dc.date.none.fl_str_mv 2018
2018-09-06
2018
2018-01-01
2018
2018-01-01
dc.type.none.fl_str_mv journal article
http://purl.org/coar/resource_type/c_6501
dc.type.openaire.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv https://hdl.handle.net/10171/53656
url https://hdl.handle.net/10171/53656
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.rights.none.fl_str_mv open access
http://purl.org/coar/access_right/c_abf2
dc.rights.openaire.fl_str_mv info:eu-repo/semantics/openAccess
rights_invalid_str_mv open access
http://purl.org/coar/access_right/c_abf2
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.source.none.fl_str_mv reponame:Dadun. Depósito Académico Digital de la Universidad de Navarra
instname:Universidad de Navarra
instname_str Universidad de Navarra
reponame_str Dadun. Depósito Académico Digital de la Universidad de Navarra
collection Dadun. Depósito Académico Digital de la Universidad de Navarra
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,301603

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