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Polymorphisms in Brucella Carbonic Anhydrase II Mediate CO(2) Dependence and Fitness in vivo
Some Brucella isolates are known to require an increased concentration of CO(2) for growth, especially in the case of primary cultures obtained directly from infected animals. Moreover, the different Brucella species and biovars show a characteristic pattern of CO(2) requirement, and this trait has...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Frontiers Media S.A.
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6915039/ https://www.ncbi.nlm.nih.gov/pubmed/31921002 http://dx.doi.org/10.3389/fmicb.2019.02751 |
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author | García Lobo, Juan M. Ortiz, Yelina Gonzalez-Riancho, Candela Seoane, Asunción Arellano-Reynoso, Beatriz Sangari, Félix J. |
author_facet | García Lobo, Juan M. Ortiz, Yelina Gonzalez-Riancho, Candela Seoane, Asunción Arellano-Reynoso, Beatriz Sangari, Félix J. |
author_sort | García Lobo, Juan M. |
collection | PubMed |
description | Some Brucella isolates are known to require an increased concentration of CO(2) for growth, especially in the case of primary cultures obtained directly from infected animals. Moreover, the different Brucella species and biovars show a characteristic pattern of CO(2) requirement, and this trait has been included among the routine typing tests used for species and biovar differentiation. By comparing the differences in gene content among different CO(2)-dependent and CO(2)-independent Brucella strains, we have confirmed that carbonic anhydrase (CA) II is the enzyme responsible for this phenotype in all the Brucella strains tested. Brucella species contain two CAs of the β family, CA I and CA II; genetic polymorphisms exist for both of them in different isolates, but only those putatively affecting the activity of CA II correlate with the CO(2) requirement of the corresponding isolate. Analysis of these polymorphisms does not allow the determination of CA I functionality, while the polymorphisms in CA II consist of small deletions that cause a frameshift that changes the C-terminus of the protein, probably affecting its dimerization status, essential for the activity. CO(2)-independent mutants arise easily in vitro, although with a low frequency ranging from 10(–6) to 10(–10) depending on the strain. These mutants carry compensatory mutations that produce a full-length CA II. At the same time, no change was observed in the sequence coding for CA I. A competitive index assay designed to evaluate the fitness of a CO(2)-dependent strain compared to its corresponding CO(2)-independent strain revealed that while there is no significant difference when the bacteria are grown in culture plates, growth in vivo in a mouse model of infection provides a significant advantage to the CO(2)-dependent strain. This could explain why some Brucella isolates are CO(2) dependent in primary isolation. The polymorphism described here also allows the in silico determination of the CO(2) requirement status of any Brucella strain. |
format | Online Article Text |
id | pubmed-6915039 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-69150392020-01-09 Polymorphisms in Brucella Carbonic Anhydrase II Mediate CO(2) Dependence and Fitness in vivo García Lobo, Juan M. Ortiz, Yelina Gonzalez-Riancho, Candela Seoane, Asunción Arellano-Reynoso, Beatriz Sangari, Félix J. Front Microbiol Microbiology Some Brucella isolates are known to require an increased concentration of CO(2) for growth, especially in the case of primary cultures obtained directly from infected animals. Moreover, the different Brucella species and biovars show a characteristic pattern of CO(2) requirement, and this trait has been included among the routine typing tests used for species and biovar differentiation. By comparing the differences in gene content among different CO(2)-dependent and CO(2)-independent Brucella strains, we have confirmed that carbonic anhydrase (CA) II is the enzyme responsible for this phenotype in all the Brucella strains tested. Brucella species contain two CAs of the β family, CA I and CA II; genetic polymorphisms exist for both of them in different isolates, but only those putatively affecting the activity of CA II correlate with the CO(2) requirement of the corresponding isolate. Analysis of these polymorphisms does not allow the determination of CA I functionality, while the polymorphisms in CA II consist of small deletions that cause a frameshift that changes the C-terminus of the protein, probably affecting its dimerization status, essential for the activity. CO(2)-independent mutants arise easily in vitro, although with a low frequency ranging from 10(–6) to 10(–10) depending on the strain. These mutants carry compensatory mutations that produce a full-length CA II. At the same time, no change was observed in the sequence coding for CA I. A competitive index assay designed to evaluate the fitness of a CO(2)-dependent strain compared to its corresponding CO(2)-independent strain revealed that while there is no significant difference when the bacteria are grown in culture plates, growth in vivo in a mouse model of infection provides a significant advantage to the CO(2)-dependent strain. This could explain why some Brucella isolates are CO(2) dependent in primary isolation. The polymorphism described here also allows the in silico determination of the CO(2) requirement status of any Brucella strain. Frontiers Media S.A. 2019-12-10 /pmc/articles/PMC6915039/ /pubmed/31921002 http://dx.doi.org/10.3389/fmicb.2019.02751 Text en Copyright © 2019 García Lobo, Ortiz, González-Riancho, Seoane, Arellano-Reynoso and Sangari. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Microbiology García Lobo, Juan M. Ortiz, Yelina Gonzalez-Riancho, Candela Seoane, Asunción Arellano-Reynoso, Beatriz Sangari, Félix J. Polymorphisms in Brucella Carbonic Anhydrase II Mediate CO(2) Dependence and Fitness in vivo |
title | Polymorphisms in Brucella Carbonic Anhydrase II Mediate CO(2) Dependence and Fitness in vivo |
title_full | Polymorphisms in Brucella Carbonic Anhydrase II Mediate CO(2) Dependence and Fitness in vivo |
title_fullStr | Polymorphisms in Brucella Carbonic Anhydrase II Mediate CO(2) Dependence and Fitness in vivo |
title_full_unstemmed | Polymorphisms in Brucella Carbonic Anhydrase II Mediate CO(2) Dependence and Fitness in vivo |
title_short | Polymorphisms in Brucella Carbonic Anhydrase II Mediate CO(2) Dependence and Fitness in vivo |
title_sort | polymorphisms in brucella carbonic anhydrase ii mediate co(2) dependence and fitness in vivo |
topic | Microbiology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6915039/ https://www.ncbi.nlm.nih.gov/pubmed/31921002 http://dx.doi.org/10.3389/fmicb.2019.02751 |
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