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Transcription Analysis of Central Metabolism Genes in Escherichia coli. Possible Roles of σ(38) in Their Expression, as a Response to Carbon Limitation

The phosphoenolpyruvate: carbohydrate transferase system (PTS) transports glucose in Escherichia coli. Previous work demonstrated that strains lacking PTS, such as PB11, grow slow on glucose. PB11 has a reduced expression of glycolytic, and upregulates poxB and acs genes as compared to the parental...

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Autores principales: Olvera, Leticia, Mendoza-Vargas, Alfredo, Flores, Noemí, Olvera, Maricela, Sigala, Juan Carlos, Gosset, Guillermo, Morett, Enrique, Bolívar, Francisco
Formato: Texto
Lenguaje:English
Publicado: Public Library of Science 2009
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2759082/
https://www.ncbi.nlm.nih.gov/pubmed/19838295
http://dx.doi.org/10.1371/journal.pone.0007466
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author Olvera, Leticia
Mendoza-Vargas, Alfredo
Flores, Noemí
Olvera, Maricela
Sigala, Juan Carlos
Gosset, Guillermo
Morett, Enrique
Bolívar, Francisco
author_facet Olvera, Leticia
Mendoza-Vargas, Alfredo
Flores, Noemí
Olvera, Maricela
Sigala, Juan Carlos
Gosset, Guillermo
Morett, Enrique
Bolívar, Francisco
author_sort Olvera, Leticia
collection PubMed
description The phosphoenolpyruvate: carbohydrate transferase system (PTS) transports glucose in Escherichia coli. Previous work demonstrated that strains lacking PTS, such as PB11, grow slow on glucose. PB11 has a reduced expression of glycolytic, and upregulates poxB and acs genes as compared to the parental strain JM101, when growing on glucose. The products of the latter genes are involved in the production of AcetylCoA. Inactivation of rpoS that codes for the RNA polymerase σ(38) subunit, reduces further (50%) growth of PB11, indicating that σ(38) plays a central role in the expression of central metabolism genes in slowly growing cells. In fact, transcription levels of glycolytic genes is reduced in strain PB11rpoS (−) as compared to PB11. In this report we studied the role of σ(70) and σ(38) in the expression of the complete glycolytic pathway and poxB and acs genes in certain PTS(−) strains and their rpoS (−) derivatives. We determined the transcription start sites (TSSs) and the corresponding promoters, in strains JM101, PB11, its derivative PB12 that recovered its growth capacity, and in their rpoS(−) derivatives, by 5′RACE and pyrosequencing. In all these genes the presence of sequences resembling σ(38) recognition sites allowed the proposition that they could be transcribed by both sigma factors, from overlapping putative promoters that initiate transcription at the same site. Fourteen new TSSs were identified in seventeen genes. Besides, more than 30 putative promoters were proposed and we confirmed ten previously reported. In vitro transcription experiments support the functionality of putative dual promoters. Alternatives that could also explain lower transcription levels of the rpoS (−) derivatives are discussed. We propose that the presence if real, of both σ(70) and σ(38) dependent promoters in all glycolytic genes and operons could allow a differential transcription of these central metabolism genes by both sigma subunits as an adaptation response to carbon limitation.
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spelling pubmed-27590822009-10-19 Transcription Analysis of Central Metabolism Genes in Escherichia coli. Possible Roles of σ(38) in Their Expression, as a Response to Carbon Limitation Olvera, Leticia Mendoza-Vargas, Alfredo Flores, Noemí Olvera, Maricela Sigala, Juan Carlos Gosset, Guillermo Morett, Enrique Bolívar, Francisco PLoS One Research Article The phosphoenolpyruvate: carbohydrate transferase system (PTS) transports glucose in Escherichia coli. Previous work demonstrated that strains lacking PTS, such as PB11, grow slow on glucose. PB11 has a reduced expression of glycolytic, and upregulates poxB and acs genes as compared to the parental strain JM101, when growing on glucose. The products of the latter genes are involved in the production of AcetylCoA. Inactivation of rpoS that codes for the RNA polymerase σ(38) subunit, reduces further (50%) growth of PB11, indicating that σ(38) plays a central role in the expression of central metabolism genes in slowly growing cells. In fact, transcription levels of glycolytic genes is reduced in strain PB11rpoS (−) as compared to PB11. In this report we studied the role of σ(70) and σ(38) in the expression of the complete glycolytic pathway and poxB and acs genes in certain PTS(−) strains and their rpoS (−) derivatives. We determined the transcription start sites (TSSs) and the corresponding promoters, in strains JM101, PB11, its derivative PB12 that recovered its growth capacity, and in their rpoS(−) derivatives, by 5′RACE and pyrosequencing. In all these genes the presence of sequences resembling σ(38) recognition sites allowed the proposition that they could be transcribed by both sigma factors, from overlapping putative promoters that initiate transcription at the same site. Fourteen new TSSs were identified in seventeen genes. Besides, more than 30 putative promoters were proposed and we confirmed ten previously reported. In vitro transcription experiments support the functionality of putative dual promoters. Alternatives that could also explain lower transcription levels of the rpoS (−) derivatives are discussed. We propose that the presence if real, of both σ(70) and σ(38) dependent promoters in all glycolytic genes and operons could allow a differential transcription of these central metabolism genes by both sigma subunits as an adaptation response to carbon limitation. Public Library of Science 2009-10-19 /pmc/articles/PMC2759082/ /pubmed/19838295 http://dx.doi.org/10.1371/journal.pone.0007466 Text en Olvera et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Olvera, Leticia
Mendoza-Vargas, Alfredo
Flores, Noemí
Olvera, Maricela
Sigala, Juan Carlos
Gosset, Guillermo
Morett, Enrique
Bolívar, Francisco
Transcription Analysis of Central Metabolism Genes in Escherichia coli. Possible Roles of σ(38) in Their Expression, as a Response to Carbon Limitation
title Transcription Analysis of Central Metabolism Genes in Escherichia coli. Possible Roles of σ(38) in Their Expression, as a Response to Carbon Limitation
title_full Transcription Analysis of Central Metabolism Genes in Escherichia coli. Possible Roles of σ(38) in Their Expression, as a Response to Carbon Limitation
title_fullStr Transcription Analysis of Central Metabolism Genes in Escherichia coli. Possible Roles of σ(38) in Their Expression, as a Response to Carbon Limitation
title_full_unstemmed Transcription Analysis of Central Metabolism Genes in Escherichia coli. Possible Roles of σ(38) in Their Expression, as a Response to Carbon Limitation
title_short Transcription Analysis of Central Metabolism Genes in Escherichia coli. Possible Roles of σ(38) in Their Expression, as a Response to Carbon Limitation
title_sort transcription analysis of central metabolism genes in escherichia coli. possible roles of σ(38) in their expression, as a response to carbon limitation
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2759082/
https://www.ncbi.nlm.nih.gov/pubmed/19838295
http://dx.doi.org/10.1371/journal.pone.0007466
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