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Functional characterization and proteomic analysis of lolA in Xanthomonas campestris pv. campestris
BACKGROUND: The gram-negative Xanthomonas campestris pv. campestris is the pathogenic bacterium that causes black rot disease in crucifers. The virulence determinants of this bacterium include extracellular enzymes, exopolysaccharides, and biofilm formation. Here, one transposon mutant of X. campest...
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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BioMed Central
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6341742/ https://www.ncbi.nlm.nih.gov/pubmed/30665348 http://dx.doi.org/10.1186/s12866-019-1387-9 |
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author | Liao, Chao-Tsai Chiang, Ying-Chuan Hsiao, Yi-Min |
author_facet | Liao, Chao-Tsai Chiang, Ying-Chuan Hsiao, Yi-Min |
author_sort | Liao, Chao-Tsai |
collection | PubMed |
description | BACKGROUND: The gram-negative Xanthomonas campestris pv. campestris is the pathogenic bacterium that causes black rot disease in crucifers. The virulence determinants of this bacterium include extracellular enzymes, exopolysaccharides, and biofilm formation. Here, one transposon mutant of X. campestris pv. campestris strain 17 that affects biofilm formation was isolated, and subsequent analyses led to the identification of the lolA gene, which encodes an outer membrane lipoprotein chaperone. RESULTS: The lolA mutant exhibited significant reductions in bacterial attachment, extracellular enzyme production, virulence, and tolerance in the presence of myriad membrane-perturbing agents. These phenotypic changes of the mutant could be complemented to the wild-type level through the intact lolA gene. Proteomic analysis revealed that 109 proteins were differentially expressed after lolA mutation. These differentially expressed proteins were categorized in various functional groups and were mainly associated with the membrane component, were involved in transport, and contained receptor activity. Through reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) analysis, deletion of lolA was determined to have caused significantly reduced expression of genes that encode the major extracellular enzymes, the biofilm-related proteins, and the virulence-related proteins. The RT-qPCR analysis also indicated that the expression of several genes that encode putative outer membrane lipoproteins and TonB-dependent receptors was reduced after lolA mutation. CONCLUSIONS: This is the first report to define the lolA gene as a virulence factor and to contribute to the functional understanding of, and provide new information concerning, the role of lolA in Xanthomonas. Furthermore, the results of this study provide and extend new insights into the function of lolA in bacteria. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12866-019-1387-9) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-6341742 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-63417422019-01-24 Functional characterization and proteomic analysis of lolA in Xanthomonas campestris pv. campestris Liao, Chao-Tsai Chiang, Ying-Chuan Hsiao, Yi-Min BMC Microbiol Research Article BACKGROUND: The gram-negative Xanthomonas campestris pv. campestris is the pathogenic bacterium that causes black rot disease in crucifers. The virulence determinants of this bacterium include extracellular enzymes, exopolysaccharides, and biofilm formation. Here, one transposon mutant of X. campestris pv. campestris strain 17 that affects biofilm formation was isolated, and subsequent analyses led to the identification of the lolA gene, which encodes an outer membrane lipoprotein chaperone. RESULTS: The lolA mutant exhibited significant reductions in bacterial attachment, extracellular enzyme production, virulence, and tolerance in the presence of myriad membrane-perturbing agents. These phenotypic changes of the mutant could be complemented to the wild-type level through the intact lolA gene. Proteomic analysis revealed that 109 proteins were differentially expressed after lolA mutation. These differentially expressed proteins were categorized in various functional groups and were mainly associated with the membrane component, were involved in transport, and contained receptor activity. Through reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) analysis, deletion of lolA was determined to have caused significantly reduced expression of genes that encode the major extracellular enzymes, the biofilm-related proteins, and the virulence-related proteins. The RT-qPCR analysis also indicated that the expression of several genes that encode putative outer membrane lipoproteins and TonB-dependent receptors was reduced after lolA mutation. CONCLUSIONS: This is the first report to define the lolA gene as a virulence factor and to contribute to the functional understanding of, and provide new information concerning, the role of lolA in Xanthomonas. Furthermore, the results of this study provide and extend new insights into the function of lolA in bacteria. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12866-019-1387-9) contains supplementary material, which is available to authorized users. BioMed Central 2019-01-21 /pmc/articles/PMC6341742/ /pubmed/30665348 http://dx.doi.org/10.1186/s12866-019-1387-9 Text en © The Author(s). 2019 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. 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 Liao, Chao-Tsai Chiang, Ying-Chuan Hsiao, Yi-Min Functional characterization and proteomic analysis of lolA in Xanthomonas campestris pv. campestris |
title | Functional characterization and proteomic analysis of lolA in Xanthomonas campestris pv. campestris |
title_full | Functional characterization and proteomic analysis of lolA in Xanthomonas campestris pv. campestris |
title_fullStr | Functional characterization and proteomic analysis of lolA in Xanthomonas campestris pv. campestris |
title_full_unstemmed | Functional characterization and proteomic analysis of lolA in Xanthomonas campestris pv. campestris |
title_short | Functional characterization and proteomic analysis of lolA in Xanthomonas campestris pv. campestris |
title_sort | functional characterization and proteomic analysis of lola in xanthomonas campestris pv. campestris |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6341742/ https://www.ncbi.nlm.nih.gov/pubmed/30665348 http://dx.doi.org/10.1186/s12866-019-1387-9 |
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