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Proc Natl Acad Sci U S A. 2011 Jul 26;108(30):12467-72. doi: 10.1073/pnas.1107894108. Epub 2011 Jul 12.

Mapping the regulon of Vibrio cholerae ferric uptake regulator expands its known network of gene regulation.

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1
Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, MA 02115, USA.

Abstract

ChIP coupled with next-generation sequencing (ChIP-seq) has revolutionized whole-genome mapping of DNA-binding protein sites. Although ChIP-seq rapidly gained support in eukaryotic systems, it remains underused in the mapping of bacterial transcriptional regulator-binding sites. Using the virulence-required iron-responsive ferric uptake regulator (Fur), we report a simple, broadly applicable ChIP-seq method in the pathogen Vibrio cholerae. Combining our ChIP-seq results with available microarray data, we clarify direct and indirect Fur regulation of known iron-responsive genes. We validate a subset of Fur-binding sites in vivo and show a common motif present in all Fur ChIP-seq peaks that has enhanced binding affinity for purified V. cholerae Fur. Further analysis shows that V. cholerae Fur directly regulates several additional genes associated with Fur-binding sites, expanding the role of this transcription factor into the regulation of ribosome formation, additional transport functions, and unique sRNAs.

PMID:
21750152
PMCID:
PMC3145737
DOI:
10.1073/pnas.1107894108
[Indexed for MEDLINE]
Free PMC Article
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