Organization and architecture of AggR-dependent promoters from enteroaggregative Escherichia coli

Muhammad Yasir; Christopher Icke; Radwa Abdelwahab; James R Haycocks; Rita E Godfrey; Pavelas Sazinas; Mark J Pallen; Ian R Henderson; Stephen J W Busby; Douglas F Browning

doi:10.1111/mmi.14172

Organization and architecture of AggR-dependent promoters from enteroaggregative Escherichia coli

Muhammad Yasir, Christopher Icke, Radwa Abdelwahab, James R Haycocks, Rita E Godfrey, Pavelas Sazinas, Mark J Pallen, Ian R Henderson, Stephen J W Busby, Douglas F Browning

Research output: Contribution to journal › Article › peer-review

Abstract

Enteroaggregative Escherichia coli (EAEC), is a diarrhoeagenic human pathogen commonly isolated from patients in both developing and industrialized countries. Pathogenic EAEC strains possess many virulence determinants, which are thought to be involved in causing disease, though, the exact mechanism by which EAEC causes diarrhoea is unclear. Typical EAEC strains possess the transcriptional regulator, AggR, which controls the expression of many virulence determinants, including the attachment adherence fimbriae (AAF) that are necessary for adherence to human gut epithelial cells. Here, using RNA-sequencing, we have investigated the AggR regulon from EAEC strain 042 and show that AggR regulates the transcription of genes on both the bacterial chromosome and the large virulence plasmid, pAA2. Due to the importance of fimbriae, we focused on the two AAF/II fimbrial gene clusters in EAEC 042 (afaB-aafCB and aafDA) and identified the promoter elements and AggR-binding sites required for fimbrial expression. In addition, we examined the organization of the fimbrial operon promoters from other important EAEC strains to understand the rules of AggR-dependent activation. Finally, we generated a series of semi-synthetic promoters to define the minimal sequence required for AggR-mediated activation and show that the correct positioning of a single AggR-binding site is sufficient to confer AggR-dependence.

Original language	English
Pages (from-to)	534-551
Number of pages	18
Journal	Molecular Microbiology
Volume	111
Issue number	2
Early online date	18 Dec 2018
DOIs	https://doi.org/10.1111/mmi.14172
Publication status	Published - Feb 2019

Bibliographical note

© 2018 The Authors. Molecular Microbiology Published by John Wiley & Sons Ltd

This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

Funding: Darwin Trust of Edinburgh PhD studentship; Industrial Biotechnology Catalyst (Innovate UK, BBSRC, EPSRC)

Keywords

Binding Sites
DNA, Bacterial/genetics
Escherichia coli/genetics
Escherichia coli Proteins/metabolism
Gene Expression Profiling
Gene Expression Regulation, Bacterial
Promoter Regions, Genetic
Protein Binding
Regulon
Sequence Analysis, RNA
Trans-Activators/metabolism

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10.1111/mmi.14172Licence: CC BY 3.0

Organization and architecture of AggR‐dependent promoters from enteroaggregative
© 2018 The Authors. Molecular Microbiology Published by John Wiley & Sons Ltd This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
Final published version, 1.02 MBLicence: CC BY 3.0

Cite this

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title = "Organization and architecture of AggR-dependent promoters from enteroaggregative Escherichia coli",

abstract = "Enteroaggregative Escherichia coli (EAEC), is a diarrhoeagenic human pathogen commonly isolated from patients in both developing and industrialized countries. Pathogenic EAEC strains possess many virulence determinants, which are thought to be involved in causing disease, though, the exact mechanism by which EAEC causes diarrhoea is unclear. Typical EAEC strains possess the transcriptional regulator, AggR, which controls the expression of many virulence determinants, including the attachment adherence fimbriae (AAF) that are necessary for adherence to human gut epithelial cells. Here, using RNA-sequencing, we have investigated the AggR regulon from EAEC strain 042 and show that AggR regulates the transcription of genes on both the bacterial chromosome and the large virulence plasmid, pAA2. Due to the importance of fimbriae, we focused on the two AAF/II fimbrial gene clusters in EAEC 042 (afaB-aafCB and aafDA) and identified the promoter elements and AggR-binding sites required for fimbrial expression. In addition, we examined the organization of the fimbrial operon promoters from other important EAEC strains to understand the rules of AggR-dependent activation. Finally, we generated a series of semi-synthetic promoters to define the minimal sequence required for AggR-mediated activation and show that the correct positioning of a single AggR-binding site is sufficient to confer AggR-dependence.",

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AU - Icke, Christopher

AU - Abdelwahab, Radwa

AU - Haycocks, James R

AU - Godfrey, Rita E

AU - Sazinas, Pavelas

AU - Pallen, Mark J

AU - Henderson, Ian R

AU - Busby, Stephen J W

AU - Browning, Douglas F

N1 - © 2018 The Authors. Molecular Microbiology Published by John Wiley & Sons Ltd This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. Funding: Darwin Trust of Edinburgh PhD studentship; Industrial Biotechnology Catalyst (Innovate UK, BBSRC, EPSRC)

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N2 - Enteroaggregative Escherichia coli (EAEC), is a diarrhoeagenic human pathogen commonly isolated from patients in both developing and industrialized countries. Pathogenic EAEC strains possess many virulence determinants, which are thought to be involved in causing disease, though, the exact mechanism by which EAEC causes diarrhoea is unclear. Typical EAEC strains possess the transcriptional regulator, AggR, which controls the expression of many virulence determinants, including the attachment adherence fimbriae (AAF) that are necessary for adherence to human gut epithelial cells. Here, using RNA-sequencing, we have investigated the AggR regulon from EAEC strain 042 and show that AggR regulates the transcription of genes on both the bacterial chromosome and the large virulence plasmid, pAA2. Due to the importance of fimbriae, we focused on the two AAF/II fimbrial gene clusters in EAEC 042 (afaB-aafCB and aafDA) and identified the promoter elements and AggR-binding sites required for fimbrial expression. In addition, we examined the organization of the fimbrial operon promoters from other important EAEC strains to understand the rules of AggR-dependent activation. Finally, we generated a series of semi-synthetic promoters to define the minimal sequence required for AggR-mediated activation and show that the correct positioning of a single AggR-binding site is sufficient to confer AggR-dependence.

AB - Enteroaggregative Escherichia coli (EAEC), is a diarrhoeagenic human pathogen commonly isolated from patients in both developing and industrialized countries. Pathogenic EAEC strains possess many virulence determinants, which are thought to be involved in causing disease, though, the exact mechanism by which EAEC causes diarrhoea is unclear. Typical EAEC strains possess the transcriptional regulator, AggR, which controls the expression of many virulence determinants, including the attachment adherence fimbriae (AAF) that are necessary for adherence to human gut epithelial cells. Here, using RNA-sequencing, we have investigated the AggR regulon from EAEC strain 042 and show that AggR regulates the transcription of genes on both the bacterial chromosome and the large virulence plasmid, pAA2. Due to the importance of fimbriae, we focused on the two AAF/II fimbrial gene clusters in EAEC 042 (afaB-aafCB and aafDA) and identified the promoter elements and AggR-binding sites required for fimbrial expression. In addition, we examined the organization of the fimbrial operon promoters from other important EAEC strains to understand the rules of AggR-dependent activation. Finally, we generated a series of semi-synthetic promoters to define the minimal sequence required for AggR-mediated activation and show that the correct positioning of a single AggR-binding site is sufficient to confer AggR-dependence.

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KW - DNA, Bacterial/genetics

KW - Escherichia coli/genetics

KW - Escherichia coli Proteins/metabolism

KW - Gene Expression Profiling

KW - Gene Expression Regulation, Bacterial

KW - Promoter Regions, Genetic

KW - Protein Binding

KW - Regulon

KW - Sequence Analysis, RNA

KW - Trans-Activators/metabolism

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DO - 10.1111/mmi.14172

M3 - Article

C2 - 30485564

SN - 0950-382X

VL - 111

SP - 534

EP - 551

JO - Molecular Microbiology

JF - Molecular Microbiology

IS - 2

ER -

Organization and architecture of AggR-dependent promoters from enteroaggregative Escherichia coli

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Keywords

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