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Reconstruction of a Pan-regulon for Fur uncovers the conservation of its transcriptional regulation in E. coli

ABSTRACT: Regulons for many transcription factors have been elucidated in model strains leading to an understanding of their role in producing physiological states. Comparative analysis of a regulon and its target genes between different strains of the same species is lacking. Ferric uptake regulator (Fur), involved in iron homeostasis, is one of the most conserved TFs, and is present in a wide range of bacteria. Using ChIP-exo experiments, we performed a comprehensive study of Fur binding sites in nine Escherichia coli strains with different lifestyles. 79 of the 431 target genes (18%) found belong to Fur’s core regulon, comprising genes involved in ion transport and metabolism, energy production and conversion, and amino acid metabolism and transport. 179 of the target genes (42%) comprise the accessory regulon, most of which were related to cell wall structure and biogenesis, and virulence factor pathways. The remaining target genes (173 or 40%) were in the unique regulon, with gene functions that were largely unknown. Furthermore, deletion of the fur gene led to distinct phenotypes in growth, motility, antibiotic resistance, and siderophore production. These results provide a more complete understanding of how Fur regulates a set of target genes with surprising variation in closely related bacteria.

ORGANISM(S): Escherichia coli K-12 Escherichia coli Escherichia coli O157:H7 Salmonella enterica

PROVIDER: GSE150501 | GEO | 2023/08/04

REPOSITORIES: GEO

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The Fur Pan-regulon in E. coli shows diversity in strain-specific sets of target genes

Project description:Regulons for many transcription factors have been elucidated in model strains leading to an understanding of their role in producing physiological states. Comparative analysis of a regulon and its target genes between different strains of the same species is lacking. Ferric uptake regulator (Fur), involved in iron homeostasis, is one of the most conserved TFs, and is present in a wide range of bacteria. Using ChIP-exo experiments, we performed a comprehensive study of Fur binding sites in nine Escherichia coli strains with different lifestyles. 79 of the 431 target genes (18%) found belong to Fur’s core regulon, comprising genes involved in ion transport and metabolism, energy production and conversion, and amino acid metabolism and transport. 179 of the target genes (42%) comprise the accessory regulon, most of which were related to cell wall structure and biogenesis, and virulence factor pathways. The remaining target genes (173 or 40%) were in the unique regulon, with gene functions that were largely unknown. Furthermore, deletion of the fur gene led to distinct phenotypes in growth, motility, antibiotic resistance, and siderophore production. These results provide a more complete understanding of how Fur regulates a set of target genes with surprising variation in closely related bacteria.

2023-08-04 | GSE150240 | GEO

The Escherichia coli Fur pan-regulon has few conserved but many unique regulatory targets

Project description:While global transcription factors (TFs) have been studied extensively in Escherichia coli model strains, conservation and diversity in TF regulation between strains is still unknown. Here we use a combination of ChIP-exo--to define ferric uptake regulator (Fur) binding sites--and differential gene expression--to define the Fur regulon in nine E. coli strains. We then define a pan-regulon consisting of 469 target genes that includes all Fur target genes in all nine strains. The pan-regulon is then divided into the core regulon (target genes found in all the strains, n=36), the accessory regulon (target found in two to eight strains, n=158) and the unique regulon (target genes found in one strain, n=275). Thus, there is a small set of Fur regulated genes common to all nine strains, but a large number of regulatory targets unique to a particular strain. Many of the unique regulatory targets are genes unique to that strain. This first-established pan-regulon reveals a common core of conserved regulatory targets and significant diversity in transcriptional regulation amongst E. coli strains, reflecting diverse niche specification and strain history.

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FUR- mutant vs. WT

Project description:Ferric uptake regulator (Fur) is the major regulator of iron acquisition in Escherichia coli and other bacteria. In the present study, the role of Fur in anaerobic S. enterica serovar Typhimurium was determined by transcriptome analysis, reporter assays, and enzymatic assays. In anaerobic Δfur, 298 genes were differentially expressed. In general, Fur repressed genes required for iron acquisition/storage, metabolism, electron transport, oxidative/nitrosative stress, and modulators of virulence. There were 73 genes whose expression required Fur, eleven of which contain a putative Fur box 5‟ of the gene. Transcription of sodA was >9-fold higher in Δfur but there was no corresponding increase in the activity of SodA.

2011-10-14 | GSE18441 | GEO

TRANSCRIPTIONAL PROFILING OF FUR– AND IRON- REGULATED GENE EXPRESSION IN LISTERIA MONOCYTOGENES

Project description:Iron is required by almost all bacteria but concentrations of iron above the physiological levels are toxic. In bacteria, intracellular iron is regulated mostly by ferric uptake regulator, Fur or a similar functional protein. Iron limitation results in regulation of number of genes, including those involved in iron uptake process. A subset of the genes under the control of Fur is called Fur regulon. In this study we have identified the Fur- and iron- regulated genes in Listeria monocytogenes by DNA microarray analysis using fur mutant and its isogenic parent. In order to identify genes exclusively regulated in response to iron limitation, fur mutant and its isogenic parent grown in iron-deficient KRM media were compared. Similarly, to identify Fur regulated genes, fur mutant was compared with the parent under iron-deficient and excess-iron conditions. Listeria in low-iron resulted in up-regulation of 180 and down-regulation of 200 genes whereas in the fur mutant, 79 genes were up-regulated and 49 genes down-regulated. Our studies have identified at least 12 genes that are negatively controlled by Fur. In iron-deficient condition, these genes were up-regulated, while expression of fur was down-regulated. To further investigate the fur regulation, the fur promoter-lacZ transcriptional fusion strains were constructed in fur and perR mutant background. In the perR mutant, the regulation of fur, svpA, and feoB was inhibited in the iron-limited conditions. Our results indicate that fur is negatively regulated by Fur and PerR. Furthermore, these results demonstrate that regulation of some of the genes in iron-limited conditions require PerR.

2008-12-25 | GSE12735 | GEO

Bacillus subtilis CU1065: WT vs fur, fur vs fur fsrA, fur vs fur fbpAB, fur vs fur fbpC, and fur vs fur fbpABC

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2011-02-19 | GSE27416 | GEO

RNA-seq analysis of gene expression in Helicobacter pylori expressing either WT Fur or a Fur variant (Fur-R88H).

Project description:Purpose: Next-generation sequencing (NGS) was used to analyze gene expression in H. pylori strains expressing either WT Fur or a Fur variant (i.e. Fur-R88H). The goals of this study are to examine the role that WT Fur and Fur-R88H play in affecting salt-responsive gene expression in the 2 different strains. The H. pylori strains were grown in the presence of routine salt (0.5% added NaCl) or high salt (1.0% added NaCl).

2023-01-06 | GSE214216 | GEO

Transcriptional analysis of the Neisseria gonorrhoeae Fur regulon

Project description:In Neisseria gonorrhoeae, Fur (ferric uptake regulator) protein regulates iron homeostasis gene expression through binding to conserved sequences in promoters of iron-responsive genes. We have expanded the gonococcal Fur regulon using a custom microarray to monitor iron-responsive gene expression throughout the growth curve combined with a genome-wide in silico analysis to predict Fur boxes (FB), and in vivo FuRTA assays to detect genes able to bind Fur. Keywords: time course: (1hr ,2hr, 3hr, 4hr)

2010-01-01 | GSE16352 | GEO

Porphyromonas gingivalis 33277 wild-type vs Fur orthologue Har isogenic mutant ECR455

Project description:Porphyromonas gingivalis is a major pathogen associated with the microbial biofilm-mediated disease chronic periodontitis. P. gingivalis has an obligate requirement for iron and protoporphyrin IX which it satisfies by transporting heme and iron liberated from the human host. The level of cellular iron in P. gingivalis affects the expression of a distinct iron-associated regulon of 64 genes and low iron invokes an iron sparing response. Iron homeostasis is usually mediated in Gram-negative bacteria at the transcriptional level by the Ferric Uptake Regulator (Fur). There is a single predicted P. gingivalis Fur superfamily orthologue named Har (heme associated regulator) that lacks the conserved metal binding residues found in other Fur orthologues. We show that Har binds both heme and ferrous iron resulting in a conformational change in the protein. Har was unable to complement the Escherichia coli H1780 fur mutant and there was no change in cellular metal content in a P. gingivalis Har mutant compared with the wild-type. The Har regulon of 44 genes is not predicted to play a role in iron homeostasis. Together these data indicated that Har does not regulate iron homeostasis in P. gingivalis. However, Har was required for heme-responsive biofilm development and its regulon overlapped P. gingivalis regulons previously identified after growth in heme limitation or as a homotypic biofilm. P. gingivalis is unique as an iron-dependent Gram-negative bacterium with a single heme-binding Fur superfamily orthologue, Har, that does not regulate iron homeostasis.

2015-04-01 | GSE37099 | GEO

Deciphering the Fur transcriptional regulatory network highlights its complex role beyond iron metabolism in Escherichia coli [RNA-seq]

Project description:The ferric uptake regulator (Fur) plays a critical role in the transcriptional regulation of iron metabolism in many bacteria. However, the full regulatory potential of Fur beyond iron metabolism remains undefined. Here, we comprehensively reconstructed the Fur transcriptional regulatory network in Escherichia coli K-12 MG1655 in response to iron availability using genome-wide measurements (ChIP-exo and RNA-seq). Polyomic data analysis revealed that a total of 81 genes in 42 transcription units (TUs) are directly regulated by three different modes of Fur regulation, including apo- and holo-Fur activation as well as holo-Fur repression. We showed that Fur connects iron transport and utilization enzymes with negative-feedback loop pairs for iron homeostasis. In addition, direct involvement of Fur in the regulation of DNA synthesis, energy metabolism, and biofilm development was found. These results indicate that Fur exhibits a comprehensive regulatory role affecting many fundamental cellular processes linked to iron metabolism in order to coordinate E. coli responses to the availability of iron.

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