QseA directly activates transcription of LEE1 in enterohemorrhagic Escherichia coli

Faith C. Sharp, Vanessa Sperandio

Research output: Contribution to journalArticle

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Abstract

Quorum sensing (QS) in enterohemorrhagic Escherichia coli (EHEC) regulates the expression of the locos of enterocyte effacement (LEE). The LEE contains five major operons named LEE1 through LEES. QseA was previously shown to be activated through QS and to activate the transcription of LEE1. The LEE1 operon encodes Ler, the transcription activator of all other LEE genes, and has two promoters: a distal promoter (P1) and a proximal promoter (P2). We have previously reported that QseA acts on P1 and not P2. To identify the minimal region of LEE1 that is necessary for QseA-mediated activation, a series of nested-deletion constructs of the LEE1 promoter fused to a lacZ reporter were constructed in both the EIIEC and E. coli K-12 backgrounds. In an EHEC background, QseA-dependent activation of LEE1 can be observed for the entire regulatory region (beginning at nucleotide -393 and ending at nucleotide -123). In contrast to what occurred in EIIEC, in K-12 there was no QseA-dependent activation of LEE1 transcription between base pairs -393 and -343. These data indicate that a QseA-dependent EHEC-specific regulator is required for the activation of transcription in this region. We also observed QseA-dependent LEE1 activation from nucleotides -218 to -123 in K-12, similar to results of the nested-deletion analysis performed with EHEC. Electrophoretic mobility shift assays established that QseA directly binds to the region of LEE1 from bp -173 to -42 and not to the region from bp -393 to -343. These studies suggest that QseA activates the transcription of LEE1 by directly binding upstream of its P1 promoter region.

Original languageEnglish (US)
Pages (from-to)2432-2440
Number of pages9
JournalInfection and Immunity
Volume75
Issue number5
DOIs
StatePublished - May 2007

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Enterohemorrhagic Escherichia coli
Enterocytes
Quorum Sensing
Nucleotides
Operon
Transcriptional Activation
Nucleic Acid Regulatory Sequences
Electrophoretic Mobility Shift Assay
Genetic Promoter Regions
Base Pairing
Escherichia coli
Genes

ASJC Scopus subject areas

  • Immunology

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QseA directly activates transcription of LEE1 in enterohemorrhagic Escherichia coli. / Sharp, Faith C.; Sperandio, Vanessa.

In: Infection and Immunity, Vol. 75, No. 5, 05.2007, p. 2432-2440.

Research output: Contribution to journalArticle

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