ClbR Is the Key Transcriptional Activator of Colibactin Gene Expression in Escherichia coli.

MSphere
Alexander WallensteinUlrich Dobrindt

Abstract

Colibactin is a nonribosomal peptide/polyketide hybrid natural product expressed by different members of the Enterobacteriaceae which can be correlated with induction of DNA double-strand breaks and interference with cell cycle progression in eukaryotes. Regulatory features of colibactin expression are only incompletely understood. We used Escherichia coli strain M1/5 as a model to investigate regulation of expression of the colibactin determinant at the transcriptional level and to characterize regulatory elements located within the colibactin pathogenicity island itself. We measured clbR transcription in vitro and observed that cultivation in defined minimal media led to increased colibactin expression relative to rich media. Transcription of clbR directly responds to iron availability. We also characterized structural DNA elements inside the colibactin determinant involved in ClbR-dependent regulation, i.e., ClbR binding sites and a variable number of tandem repeats located upstream of clbR We investigated the impact of clbR overexpression or deletion at the transcriptome and proteome levels. Moreover, we compared global gene regulation under these conditions with that occurring upon overexpression or deletion of clbQ, which...Continue Reading

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Datasets Mentioned

BETA
KUN4843
CP053296
CP053298

Methods Mentioned

BETA
biopsy
flow cytometry
electrophoretic mobility shift
RNA-seq
reverse transcription-PCR
Illumina sequencing
genetic modifications
PCR
electrophoresis

Software Mentioned

PGAP
GraphPad Prism
Clustal Omega
SerotypeFinder
SMRT
BWA
ResFinder
ReadXplorer
DESeq2
PlasmidFinder

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