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Transcriptomic analysis of the stationary phase response regulator SpdR in Caulobacter crescentus

BACKGROUND: As bacterial cells enter stationary phase, they adjust their growth rate to comply with nutrient restriction and acquire increased resistance to several stresses. These events are regulated by controlling gene expression at this phase, changing the mode of exponential growth into that of...

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Autores principales: da Silva, Carolina A. P. T., Lourenço, Rogério F., Mazzon, Ricardo R., Ribeiro, Rodolfo A., Marques, Marilis V.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4830024/
https://www.ncbi.nlm.nih.gov/pubmed/27072651
http://dx.doi.org/10.1186/s12866-016-0682-y
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author da Silva, Carolina A. P. T.
Lourenço, Rogério F.
Mazzon, Ricardo R.
Ribeiro, Rodolfo A.
Marques, Marilis V.
author_facet da Silva, Carolina A. P. T.
Lourenço, Rogério F.
Mazzon, Ricardo R.
Ribeiro, Rodolfo A.
Marques, Marilis V.
author_sort da Silva, Carolina A. P. T.
collection PubMed
description BACKGROUND: As bacterial cells enter stationary phase, they adjust their growth rate to comply with nutrient restriction and acquire increased resistance to several stresses. These events are regulated by controlling gene expression at this phase, changing the mode of exponential growth into that of growth arrest, and increasing the expression of proteins involved in stress resistance. The two-component system SpdR/SpdS is required for the activation of transcription of the Caulobacter crescentus cspD gene at the onset of stationary phase. RESULTS: In this work, we showed that both SpdR and SpdS are also induced upon entry into stationary phase, and this induction is partly mediated by ppGpp and it is not auto-regulated. Global transcriptional analysis at early stationary phase of a spdR null mutant strain compared to the wild type strain was carried out by DNA microarray. Twenty-three genes showed at least twofold decreased expression in the spdR deletion mutant strain relative to its parental strain, including cspD, while five genes showed increased expression in the mutant. The expression of a set of nine genes was evaluated by quantitative real time PCR, validating the microarray data, and indicating an important role for SpdR at stationary phase. Several of the differentially expressed genes can be involved in modulating gene expression, including four transcriptional regulators, and the RNA regulatory protein Hfq. The ribosomal proteins NusE and NusG, which also have additional regulatory functions in transcription and translation, were also downregulated in the spdR mutant, as well as the ParE1 toxin. The purified SpdR protein was shown to bind to the regulatory region of CC0517 by Electrophoretic Mobility Shift Assay, and the SpdR-regulated gene CC0731 was shown to be expressed at a lower level in the null cspD mutant, suggesting that at least part of the effect of SpdR on the expression of this gene is indirect. CONCLUSIONS: The results indicate that SpdR regulates several genes encoding proteins of regulatory function, which in turn may be required for the expression of other genes important for the transition to stationary phase. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12866-016-0682-y) contains supplementary material, which is available to authorized users.
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spelling pubmed-48300242016-04-14 Transcriptomic analysis of the stationary phase response regulator SpdR in Caulobacter crescentus da Silva, Carolina A. P. T. Lourenço, Rogério F. Mazzon, Ricardo R. Ribeiro, Rodolfo A. Marques, Marilis V. BMC Microbiol Research Article BACKGROUND: As bacterial cells enter stationary phase, they adjust their growth rate to comply with nutrient restriction and acquire increased resistance to several stresses. These events are regulated by controlling gene expression at this phase, changing the mode of exponential growth into that of growth arrest, and increasing the expression of proteins involved in stress resistance. The two-component system SpdR/SpdS is required for the activation of transcription of the Caulobacter crescentus cspD gene at the onset of stationary phase. RESULTS: In this work, we showed that both SpdR and SpdS are also induced upon entry into stationary phase, and this induction is partly mediated by ppGpp and it is not auto-regulated. Global transcriptional analysis at early stationary phase of a spdR null mutant strain compared to the wild type strain was carried out by DNA microarray. Twenty-three genes showed at least twofold decreased expression in the spdR deletion mutant strain relative to its parental strain, including cspD, while five genes showed increased expression in the mutant. The expression of a set of nine genes was evaluated by quantitative real time PCR, validating the microarray data, and indicating an important role for SpdR at stationary phase. Several of the differentially expressed genes can be involved in modulating gene expression, including four transcriptional regulators, and the RNA regulatory protein Hfq. The ribosomal proteins NusE and NusG, which also have additional regulatory functions in transcription and translation, were also downregulated in the spdR mutant, as well as the ParE1 toxin. The purified SpdR protein was shown to bind to the regulatory region of CC0517 by Electrophoretic Mobility Shift Assay, and the SpdR-regulated gene CC0731 was shown to be expressed at a lower level in the null cspD mutant, suggesting that at least part of the effect of SpdR on the expression of this gene is indirect. CONCLUSIONS: The results indicate that SpdR regulates several genes encoding proteins of regulatory function, which in turn may be required for the expression of other genes important for the transition to stationary phase. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12866-016-0682-y) contains supplementary material, which is available to authorized users. BioMed Central 2016-04-12 /pmc/articles/PMC4830024/ /pubmed/27072651 http://dx.doi.org/10.1186/s12866-016-0682-y Text en © da Silva et al. 2016 Open AccessThis 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
da Silva, Carolina A. P. T.
Lourenço, Rogério F.
Mazzon, Ricardo R.
Ribeiro, Rodolfo A.
Marques, Marilis V.
Transcriptomic analysis of the stationary phase response regulator SpdR in Caulobacter crescentus
title Transcriptomic analysis of the stationary phase response regulator SpdR in Caulobacter crescentus
title_full Transcriptomic analysis of the stationary phase response regulator SpdR in Caulobacter crescentus
title_fullStr Transcriptomic analysis of the stationary phase response regulator SpdR in Caulobacter crescentus
title_full_unstemmed Transcriptomic analysis of the stationary phase response regulator SpdR in Caulobacter crescentus
title_short Transcriptomic analysis of the stationary phase response regulator SpdR in Caulobacter crescentus
title_sort transcriptomic analysis of the stationary phase response regulator spdr in caulobacter crescentus
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4830024/
https://www.ncbi.nlm.nih.gov/pubmed/27072651
http://dx.doi.org/10.1186/s12866-016-0682-y
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