Pet is a cytotoxic autotransporter protein secreted by the pathogenic enteroaggregative Escherichia coli strain 042. Expression of Pet is co-dependent on two global transcription regulators: CRP (cyclic AMP receptor protein) and Fis (factor for inversion stimulation). At the pet promoter CRP binds to a single site centred at position -40.5 upstream of the start site for transcription. Due to the suboptimal positioning of this site, CRP alone activates transcription poorly and requires Fis to bind upstream to promote full activation. Here, we show that CRP and Fis control the expression of other important autotransporter toxins, namely Sat from uropathogenic E. coli (UPEC) and SigA from Shigella sonnei, and that this regulation has been conserved in different pathogens. Furthermore, we investigate the mechanism of Fis-mediated co-activation, exploiting a series of semi-synthetic promoters, with similar architecture to the pet promoter. We show that, when bound at position -40.5, CRP recruits RNA polymerase inefficiently and that Fis compensates by aiding polymerase recruitment through a direct protein–protein interaction. We demonstrate that other suitably positioned upstream transcription factors, which directly recruit RNA polymerase, can also compensate for the inappropriate positioning of CRP. We propose that this is a simple ‘shared-recruitment’ mechanism, by which co-dependence of promoters on two transcription factors could evolve.
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Research Article|
February 20 2015
Expression of different bacterial cytotoxins is controlled by two global transcription factors, CRP and Fis, that co-operate in a shared-recruitment mechanism Available to Purchase
Amanda E. Rossiter;
*Institute of Microbiology and Infection, University of Birmingham, Edgbaston, Birmingham B15 2TT, U.K.
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Rita E. Godfrey;
Rita E. Godfrey
1
*Institute of Microbiology and Infection, University of Birmingham, Edgbaston, Birmingham B15 2TT, U.K.
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Jack A. Connolly;
Jack A. Connolly
*Institute of Microbiology and Infection, University of Birmingham, Edgbaston, Birmingham B15 2TT, U.K.
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Stephen J. W. Busby;
Stephen J. W. Busby
*Institute of Microbiology and Infection, University of Birmingham, Edgbaston, Birmingham B15 2TT, U.K.
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Ian R. Henderson;
Ian R. Henderson
*Institute of Microbiology and Infection, University of Birmingham, Edgbaston, Birmingham B15 2TT, U.K.
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Douglas F. Browning
Douglas F. Browning
2
*Institute of Microbiology and Infection, University of Birmingham, Edgbaston, Birmingham B15 2TT, U.K.
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Publisher: Portland Press Ltd
Received:
October 23 2014
Revision Received:
December 02 2014
Accepted:
December 08 2014
Accepted Manuscript online:
December 08 2014
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© The Authors Journal compilation © 2015 Biochemical Society
2015
Biochem J (2015) 466 (2): 323–335.
Article history
Received:
October 23 2014
Revision Received:
December 02 2014
Accepted:
December 08 2014
Accepted Manuscript online:
December 08 2014
Citation
Amanda E. Rossiter, Rita E. Godfrey, Jack A. Connolly, Stephen J. W. Busby, Ian R. Henderson, Douglas F. Browning; Expression of different bacterial cytotoxins is controlled by two global transcription factors, CRP and Fis, that co-operate in a shared-recruitment mechanism. Biochem J 1 March 2015; 466 (2): 323–335. doi: https://doi.org/10.1042/BJ20141315
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