Publications by authors named "Patricia J Kiley"

The widespread family of Rrf2 transcription factors has emerged as having prominent roles in diverse bacterial functions. These proteins share an overall common structure to sense and respond to stress signals. In many known cases, signaling occurs through iron-sulfur cluster cofactors.

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Unlabelled: Isoprenoids are a diverse family of compounds that are synthesized from two isomeric compounds, isopentenyl diphosphate and dimethylallyl diphosphate. In most bacteria, isoprenoids are produced from the essential methylerythritol phosphate (MEP) pathway. The terminal enzymes of the MEP pathway IspG and IspH are [4Fe-4S] cluster proteins, and in the substrates of IspG and IspH accumulate in cells in response to O, suggesting possible lability of their [4Fe-4S] clusters.

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The role of IscR in regulating the transcription of genes involved in Fe-S cluster homeostasis has been well established for the model organism Escherichia coli K12. In this bacterium, IscR coordinates expression of the Isc and Suf Fe-S cluster assembly pathways to meet cellular Fe-S cluster demands shaped by a variety of environmental cues. However, since its initial discovery nearly 25 years ago, there has been growing evidence that IscR function extends well beyond Fe-S cluster homeostasis, not only in E.

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Myxococcus xanthus possesses two Fe-S cluster biogenesis machineries, ISC (iron-sulfur cluster) and SUF (sulfur mobilization). Here, we show that in comparison to the phylogenetically distant Enterobacteria, which also have both machineries, M. xanthus evolved an independent transcriptional scheme to coordinately regulate the expression of these machineries.

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The type III secretion system (T3SS) is an appendage used by many bacterial pathogens, such as pathogenic Yersinia, to subvert host defenses. However, because the T3SS is energetically costly and immunogenic, it must be tightly regulated in response to environmental cues to enable survival in the host. Here we show that expression of the Yersinia Ysc T3SS master regulator, LcrF, is orchestrated by the opposing activities of the repressive H-NS/YmoA histone-like protein complex and induction by the iron and oxygen-regulated IscR transcription factor.

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Bacterial two-component systems (TCSs) often function through the detection of an extracytoplasmic stimulus and the transduction of a signal by a transmembrane sensory histidine kinase. This kinase then initiates a series of reversible phosphorylation modifications to regulate the activity of a cognate, cytoplasmic response regulator as a transcription factor. Several TCSs have been implicated in the regulation of cell cycle dynamics, cell envelope integrity, or cell wall development in Escherichia coli and other well-studied Gram-negative model organisms.

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Article Synopsis
  • IscR is a global transcription factor in E. coli that regulates Fe-S cluster homeostasis by activating or repressing transcription at specific DNA sites.
  • It was found that IscR can recruit RNA polymerase even when the optimal -35 promoter element is absent, influencing its ability to activate or repress transcription based on the position of this element.
  • The study highlights that subtle shifts in the -35 element's location within the IscR binding site can drastically change IscR's role, revealing an evolutionary adaptation that allows flexible transcriptional regulation.
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Zymomonas mobilis has emerged as a promising candidate for production of high-value bioproducts from plant biomass. However, a major limitation in equipping Z. mobilis with novel pathways to achieve this goal is restriction of heterologous DNA.

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Article Synopsis
  • The study investigates the role of the IscR transcription factor in the virulence of Yersinia pseudotuberculosis, focusing on how it responds to iron depletion during infection.
  • Researchers utilized ChIP-Seq and RNA-Seq to identify genes regulated by IscR, discovering its regulation of genes linked to iron homeostasis, reactive oxygen species management, and the type III secretion system (T3SS).
  • Results showed that IscR helps bacteria adapt to iron-restricted environments in the host by regulating multiple pathways critical for survival and virulence, implying that IscR's functions are conserved even in closely related pathogens like Yersinia pestis.
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Genome-scale metabolic models have been utilized extensively in the study and engineering of the organisms they describe. Here we present the analysis of a published dataset from pooled transposon mutant fitness experiments as an approach for improving the accuracy and gene-reaction associations of a metabolic model for Zymomonas mobilis ZM4, an industrially relevant ethanologenic organism with extremely high glycolytic flux and low biomass yield. Gene essentiality predictions made by the draft model were compared to data from individual pooled mutant experiments to identify areas of the model requiring deeper validation.

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Pathogenicity islands and plasmids bear genes for pathogenesis of various pathotypes. Although there is a basic understanding of the contribution of these virulence factors to disease, less is known about variation in regulatory networks in determining disease phenotypes. Here, we dissected a regulatory network directed by the conserved iron homeostasis regulator, ferric uptake regulator (Fur), in uropathogenic (UPEC) strain CFT073.

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Article Synopsis
  • Yersinia pseudotuberculosis and Yersinia pestis require a type III secretion system (T3SS) to evade immune defenses and cause disease, but its expression needs careful regulation due to its link to growth arrest and immune recognition.
  • The expression of T3SS in Y. pseudotuberculosis is low in anaerobic, iron-rich conditions typical of the intestines, but increases in aerobic or iron-poor conditions faced when they encounter immune cells.
  • The transcription factor IscR regulates T3SS expression by responding to oxygen and iron levels, and its binding to the lcrF promoter is crucial for virulence; this regulatory mechanism is conserved in both Y. pseudotuberculosis and Y. pest
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Structural and spectroscopic analysis of iron-sulfur [Fe-S] cluster-containing proteins is often limited by the occupancy and yield of recombinantly produced proteins. Here we report that BL21(DE3), a strain routinely used to overproduce [Fe-S] cluster-containing proteins, has a nonfunctional Suf pathway, one of two [Fe-S] cluster biogenesis pathways. We confirmed that BL21(DE3) and commercially available derivatives carry a deletion that results in an in-frame fusion of and genes within the operon.

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Metabolic engineering of the biofuel-producing is necessary if we are to unlock the metabolic potential present in this non-model microbe. Manipulation of such organisms can be challenging because of the limited genetic tools for iterative genome modification. Here, we have developed an efficient method for generating markerless genomic deletions or additions in .

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Temperate bacteriophages are viruses that can incorporate their genomes into their bacterial hosts, existing there as prophages that refrain from killing the host cell until induced. Prophages are largely quiescent, but they can alter host phenotype through factors encoded in their genomes (often virulence factors) or by disrupting host genes as a result of integration. Here we describe another mechanism by which a prophage can modulate host phenotype.

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Currently, microbial conversion of lignocellulose-derived glucose and xylose to biofuels is hindered by the fact that most microbes (including Escherichia coli [E. coli], Saccharomyces cerevisiae, and Zymomonas mobilis) preferentially consume glucose first and consume xylose slowly after glucose is depleted in lignocellulosic hydrolysates. In this study, E.

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Article Synopsis
  • - The HmuRSTUV hemin uptake system in bacteria enables them to acquire iron from heme and hemoproteins, allowing them to thrive despite host attempts to sequester iron.
  • - The transcriptional regulation of this uptake system is complex, involving global regulators like IscR and Fur, which manage gene expression under different iron availability conditions to optimize hemin utilization.
  • - A Δ mutant strain showed reduced survival in whole blood, highlighting the importance of IscR in regulating various virulence factors that assist in bacterial growth within mammalian tissues, emphasizing the intricate relationship between heme uptake mechanisms and host defenses.
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Microbial populations can maximize fitness in dynamic environments through bet hedging, a process wherein a subpopulation assumes a phenotype not optimally adapted to the present environment but well adapted to an environment likely to be encountered. Here, we show that oxygen induces fluctuating expression of the trimethylamine oxide (TMAO) respiratory system of Escherichia coli, diversifying the cell population and enabling a bet-hedging strategy that permits growth following oxygen loss. This regulation by oxygen affects the variance in gene expression but leaves the mean unchanged.

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The ferric-uptake regulator (Fur) is an Fe-responsive transcription factor that coordinates iron homeostasis in many bacteria. Recently, we reported that expression of the Fur regulon is also impacted by O tension. Here, we show that for most of the Fur regulon, Fur binding and transcriptional repression increase under anaerobic conditions, suggesting that Fur is controlled by O availability.

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Significance: The Escherichia coli regulatory protein fumarate nitrate reduction (FNR) mediates a global transcriptional response upon O deprivation. Spanning nearly 40 years of research investigations, our understanding of how FNR senses and responds to O has considerably progressed despite a lack of structural information for most of that period. This knowledge has established the paradigm for how facultative anaerobic bacteria sense changes in O tension.

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Unlabelled: Iron, a major protein cofactor, is essential for most organisms. Despite the well-known effects of O2 on the oxidation state and solubility of iron, the impact of O2 on cellular iron homeostasis is not well understood. Here we report that in Escherichia coli K-12, the lack of O2 dramatically changes expression of genes controlled by the global regulators of iron homeostasis, the transcription factor Fur and the small RNA RyhB.

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