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PMID:19052235

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Contents

Citation

Cho, BK, Barrett, CL, Knight, EM, Park, YS and Palsson, BØ (2008) Genome-scale reconstruction of the Lrp regulatory network in Escherichia coli. Proc. Natl. Acad. Sci. U.S.A. 105:19462-7

Abstract

Broad-acting transcription factors (TFs) in bacteria form regulons. Here, we present a 4-step method to fully reconstruct the leucine-responsive protein (Lrp) regulon in Escherichia coli K-12 MG 1655 that regulates nitrogen metabolism. Step 1 is composed of obtaining high-resolution ChIP-chip data for Lrp, the RNA polymerase and expression profiles under multiple environmental conditions. We identified 138 unique and reproducible Lrp-binding regions and classified their binding state under different conditions. In the second step, the analysis of these data revealed 6 distinct regulatory modes for individual ORFs. In the third step, we used the functional assignment of the regulated ORFs to reconstruct 4 types of regulatory network motifs around the metabolites that are affected by the corresponding gene products. In the fourth step, we determined how leucine, as a signaling molecule, shifts the regulatory motifs for particular metabolites. The physiological structure that emerges shows the regulatory motifs for different amino acid fall into the traditional classification of amino acid families, thus elucidating the structure and physiological functions of the Lrp-regulon. The same procedure can be applied to other broad-acting TFs, opening the way to full bottom-up reconstruction of the transcriptional regulatory network in bacterial cells.

Links

PubMed PMC2614783 Online version:10.1073/pnas.0807227105

Keywords

Escherichia coli/genetics; Escherichia coli/metabolism; Escherichia coli Proteins/genetics; Escherichia coli Proteins/metabolism; Feedback, Physiological; Gene Expression Regulation, Bacterial; Gene Regulatory Networks; Genome, Bacterial; Genomics; Leucine/pharmacokinetics; Leucine-Responsive Regulatory Protein/genetics; Leucine-Responsive Regulatory Protein/metabolism; Nitrogen/metabolism; Oligonucleotide Array Sequence Analysis; RNA, Bacterial/genetics; Transcription, Genetic

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Annotations

Gene product Qualifier GO ID GO term name Evidence Code with/from Aspect Notes Status

Data

Strain/Organism Assay Analyte Variable Conditions Notes Links

MG1655

ChIP-chip

Lrp

Exponential phase w/ and w/o Rifampicin

Rifampicin effects are shown as subtracks

MG1655

ChIP-chip

Lrp

Stationary phase w/and w/o Rifampicin

Rifampicin effects shown as subtracks.

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