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Cysteine-Mediated Gene Expression and Characterization of the CmbR Regulon in Streptococcus pneumoniae

In this study, we investigated the transcriptomic response of Streptococcus pneumoniae D39 to cysteine. Transcriptome comparison of the D39 wild-type grown at a restricted concentration of cysteine (0.03 mM) to one grown at a high concentration of cysteine (50 mM) in chemically-defined medium (CDM)...

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Autores principales: Afzal, Muhammad, Manzoor, Irfan, Kuipers, Oscar P., Shafeeq, Sulman
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5131005/
https://www.ncbi.nlm.nih.gov/pubmed/27990139
http://dx.doi.org/10.3389/fmicb.2016.01929
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author Afzal, Muhammad
Manzoor, Irfan
Kuipers, Oscar P.
Shafeeq, Sulman
author_facet Afzal, Muhammad
Manzoor, Irfan
Kuipers, Oscar P.
Shafeeq, Sulman
author_sort Afzal, Muhammad
collection PubMed
description In this study, we investigated the transcriptomic response of Streptococcus pneumoniae D39 to cysteine. Transcriptome comparison of the D39 wild-type grown at a restricted concentration of cysteine (0.03 mM) to one grown at a high concentration of cysteine (50 mM) in chemically-defined medium (CDM) revealed elevated expression of various genes/operons, i.e., spd-0150, metQ, spd-0431, metEF, gshT, spd-0618, fhs, tcyB, metB-csd, metA, spd-1898, yvdE, and cysK, likely to be involved in the transport and utilization of cysteine and/or methionine. Microarray-based data were further confirmed by quantitative RT-PCR. Promoter lacZ-fusion studies and quantitative RT-PCR data showed that the transcriptional regulator CmbR acts as a transcriptional repressor of spd-0150, metEF, gshT, spd-0618, tcyB, metA, and yvdE, putatively involved in cysteine uptake and utilization. The operator site of CmbR in the promoter regions of CmbR-regulated genes is predicted and confirmed by mutating or deleting CmbR operator sites from the promoter regions of these genes.
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spelling pubmed-51310052016-12-16 Cysteine-Mediated Gene Expression and Characterization of the CmbR Regulon in Streptococcus pneumoniae Afzal, Muhammad Manzoor, Irfan Kuipers, Oscar P. Shafeeq, Sulman Front Microbiol Microbiology In this study, we investigated the transcriptomic response of Streptococcus pneumoniae D39 to cysteine. Transcriptome comparison of the D39 wild-type grown at a restricted concentration of cysteine (0.03 mM) to one grown at a high concentration of cysteine (50 mM) in chemically-defined medium (CDM) revealed elevated expression of various genes/operons, i.e., spd-0150, metQ, spd-0431, metEF, gshT, spd-0618, fhs, tcyB, metB-csd, metA, spd-1898, yvdE, and cysK, likely to be involved in the transport and utilization of cysteine and/or methionine. Microarray-based data were further confirmed by quantitative RT-PCR. Promoter lacZ-fusion studies and quantitative RT-PCR data showed that the transcriptional regulator CmbR acts as a transcriptional repressor of spd-0150, metEF, gshT, spd-0618, tcyB, metA, and yvdE, putatively involved in cysteine uptake and utilization. The operator site of CmbR in the promoter regions of CmbR-regulated genes is predicted and confirmed by mutating or deleting CmbR operator sites from the promoter regions of these genes. Frontiers Media S.A. 2016-12-01 /pmc/articles/PMC5131005/ /pubmed/27990139 http://dx.doi.org/10.3389/fmicb.2016.01929 Text en Copyright © 2016 Afzal, Manzoor, Kuipers and Shafeeq. 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) or licensor 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
Afzal, Muhammad
Manzoor, Irfan
Kuipers, Oscar P.
Shafeeq, Sulman
Cysteine-Mediated Gene Expression and Characterization of the CmbR Regulon in Streptococcus pneumoniae
title Cysteine-Mediated Gene Expression and Characterization of the CmbR Regulon in Streptococcus pneumoniae
title_full Cysteine-Mediated Gene Expression and Characterization of the CmbR Regulon in Streptococcus pneumoniae
title_fullStr Cysteine-Mediated Gene Expression and Characterization of the CmbR Regulon in Streptococcus pneumoniae
title_full_unstemmed Cysteine-Mediated Gene Expression and Characterization of the CmbR Regulon in Streptococcus pneumoniae
title_short Cysteine-Mediated Gene Expression and Characterization of the CmbR Regulon in Streptococcus pneumoniae
title_sort cysteine-mediated gene expression and characterization of the cmbr regulon in streptococcus pneumoniae
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5131005/
https://www.ncbi.nlm.nih.gov/pubmed/27990139
http://dx.doi.org/10.3389/fmicb.2016.01929
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