scholarly journals Regulation of rpoS Gene Expression in Pseudomonas: Involvement of a TetR Family Regulator

2001 ◽  
Vol 183 (12) ◽  
pp. 3712-3720 ◽  
Author(s):  
Milan Kojic ◽  
Vittorio Venturi
Keyword(s):  
Gene Expression ◽  
Gene Regulation ◽  
Quorum Sensing ◽  
Stationary Phase ◽  
Cell Density ◽  
Promoter Activity ◽  
Regulatory Protein ◽  
Insertion Mutant ◽  
Regulatory Cascade ◽  
A Cell

ABSTRACT The rpoS gene encodes the sigma factor which was identified in several gram-negative bacteria as a central regulator during stationary phase. rpoS gene regulation is known to respond to cell density, showing higher expression in stationary phase. For Pseudomonas aeruginosa, it has been demonstrated that the cell-density-dependent regulation response known as quorum sensing interacts with this regulatory response. Using therpoS promoter of P. putida, we identified a genomic Tn5 insertion mutant of P. putida which showed a 90% decrease in rpoSpromoter activity, resulting in less RpoS being present in a cell at stationary phase. Molecular analysis revealed that this mutant carried a Tn5 insertion in a gene, designatedpsrA (Pseudomonas sigma regulator), which codes for a protein (PsrA) of 26.3 kDa. PsrA contains a helix-turn-helix motif typical of DNA binding proteins and belongs to the TetR family of bacterial regulators. The homolog of thepsrA gene was identified in P. aeruginosa; the protein showed 90% identity to PsrA ofP. putida. ApsrA::Tn5 insertion mutant ofP. aeruginosa was constructed. In bothPseudomonas species, psrA was genetically linked to the SOS lexA repressor gene. Similar to what was observed for P. putida, a psrA null mutant of P. aeruginosa also showed a 90% reduction inrpoS promoter activity; both mutants could be complemented for rpoS promoter activity when thepsrA gene was provided in trans.psrA mutants of both Pseudomonas species lost the ability to induce rpoS expression at stationary phase, but they retained the ability to produce quorum-sensing autoinducer molecules. PsrA was demonstrated to negatively regulatepsrA gene expression in Pseudomonas and in Escherichia coli as well as to be capable of activating the rpoS promoter in E. coli. Our data suggest that PsrA is an important regulatory protein ofPseudomonas spp. involved in the regulatory cascade controlling rpoS gene regulation in response to cell density.

scholarly journals LuxS Is Required for Persistent Pneumococcal Carriage and Expression of Virulence and Biosynthesis Genes

2004 ◽  
Vol 72 (5) ◽  
pp. 2964-2975 ◽  
Author(s):  
Elizabeth A. Joyce ◽  
Amita Kawale ◽  
Stefano Censini ◽  
Charles C. Kim ◽  
Antonello Covacci ◽  
...  
Keyword(s):  
Gene Expression ◽  
Quorum Sensing ◽  
Cell Density ◽  
High Cell Density ◽  
Signaling System ◽  
Cellular Processes ◽  
Disease States ◽  
Pneumococcal Carriage ◽  
A Cell ◽  
Dependent Mechanism

ABSTRACT Streptococcus pneumoniae causes several diseases, including otitis media, pneumonia, and meningitis. Although little is known about the regulation of or how individual pneumococcal factors contribute to these disease states, there is evidence suggesting that some factors are regulated by a cell-density-dependent mechanism (quorum sensing). Quorum sensing allows bacteria to couple transcription with changes in cell density; bacteria achieve this by sensing and responding to small diffusible signaling molecules. We investigated how the LuxS signaling system impacts the biology of S. pneumoniae. An analysis of the transcriptional profiles of a serotype 2 strain and an isogenic luxS deletion strain utilizing an S. pneumoniae-specific microarray indicated that LuxS regulates gene expression involved in discrete cellular processes, including pneumolysin expression. Contrary to the paradigm for quorum sensing, we observed pronounced effects on transcription in early log phase, where gene expression was repressed in the mutant. Assessing the mutant for its ability to infect and cause disease in animals revealed a profound defect in ability to persist in the nasopharyngeal tissues. Our analysis of an S. pneumoniae transcriptome revealed a function for LuxS in gene regulation that is not dependent upon high cell density and is likely involved in the maintenance of pneumococcal load in susceptible hosts.

scholarly journals Peptide signaling without feedback in signal production operates as a true quorum sensing communication system in Bacillus subtilis

2021 ◽  
Vol 4 (1) ◽  
Author(s):  
Iztok Dogsa ◽  
Mihael Spacapan ◽  
Anna Dragoš ◽  
Tjaša Danevčič ◽  
Žiga Pandur ◽  
...  
Keyword(s):  
Gene Expression ◽  
Bacillus Subtilis ◽  
Quorum Sensing ◽  
Cell Density ◽  
Communication System ◽  
Communication Systems ◽  
Feedback Loop ◽  
Signal Molecules ◽  
Specific Cell ◽  
Sharp Transition

AbstractBacterial quorum sensing (QS) is based on signal molecules (SM), which increase in concentration with cell density. At critical SM concentration, a variety of adaptive genes sharply change their expression from basic level to maximum level. In general, this sharp transition, a hallmark of true QS, requires an SM dependent positive feedback loop, where SM enhances its own production. Some communication systems, like the peptide SM-based ComQXPA communication system of Bacillus subtilis, do not have this feedback loop and we do not understand how and if the sharp transition in gene expression is achieved. Based on experiments and mathematical modeling, we observed that the SM peptide ComX encodes the information about cell density, specific cell growth rate, and even oxygen concentration, which ensure power-law increase in SM production. This enables together with the cooperative response to SM (ComX) a sharp transition in gene expression level and this without the SM dependent feedback loop. Due to its ultra-sensitive nature, the ComQXPA can operate at SM concentrations that are 100–1000 times lower than typically found in other QS systems, thereby substantially reducing the total metabolic cost of otherwise expensive ComX peptide.

scholarly journals Optimal transcriptional regulation of cellular responses to sudden environmental shifts

2020 ◽  
Author(s):  
Daniel Schultz ◽  
Lev S. Tsimring
Keyword(s):  
Gene Expression ◽  
Gene Regulation ◽  
Cell Response ◽  
Cellular Responses ◽  
Natural Systems ◽  
Selective Pressures ◽  
Regulatory Strategies ◽  
Regulatory Strategy ◽  
A Cell ◽  
The Many

ABSTRACTCellular responses to sudden changes in their environment require prompt expression of the correct levels of the appropriate enzymes. These enzymes are typically regulated by transcription factors that sense the presence of inducers and control gene expression for the duration of the response. The specific choice of regulatory strategy depends on the characteristics of each cell response, with the pattern of gene expression dictated by parameters such as the affinity of the transcription factor to its binding sites and the strength of the promoters it regulates. Although much is known about how gene regulation determines the dynamics of cell responses, we still lack a framework to understand how the many different regulatory strategies evolved in natural systems relate to the constraints imposed by the selective pressures acting in each particular case. Here, we analyze a dynamical model of a cell response where expression of a transcriptionally repressed enzyme is induced by a sudden exposure to its substrate. We identify strategies of gene regulation that optimize the response for different types of selective pressures, which we define as a set of costs associated with substrate, enzyme and repressor intracellular concentrations during the response. We find that regulated responses happen within a defined region in the parameter space. While responses to costly (toxic) substrates favor the usage of strongly self-regulated repressors, responses where expression of enzyme is more costly than its substrate favor the usage of constitutively expressed repressors. There is only a very narrow range of selective pressures that would favor weakly self-regulated repressors. This framework can be used to infer which costs and benefits are most critical in the evolution of natural examples of cellular responses, and to predict how a response can optimize its regulation when transported to a new environment with different demands.

scholarly journals Stringent Response Activates Quorum Sensing and Modulates Cell Density-Dependent Gene Expression inPseudomonas aeruginosa

2001 ◽  
Vol 183 (18) ◽  
pp. 5376-5384 ◽  
Author(s):  
Christian van Delden ◽  
Rachel Comte ◽  
And Marc Bally
Keyword(s):  
Gene Expression ◽  
Quorum Sensing ◽  
Cell Density ◽  
Stringent Response ◽  
Transcriptional Regulators ◽  
Homoserine Lactone ◽  
Density Dependent ◽  
Sensing Systems ◽  
Acid Analogue ◽  
Enhanced Expression

ABSTRACT During nutrient starvation, Escherichia coli elicits a stringent response involving the ribosome-associated protein RelA. Activation of RelA results in a global change in the cellular metabolism including enhanced expression of the stationary-phase sigma factor RpoS. In the human pathogen Pseudomonas aeruginosa, a complex quorum-sensing circuitry, linked to RpoS expression, is required for cell density-dependent production of many secreted virulence factors, including LasB elastase. Quorum sensing relies on the activation of specific transcriptional regulators (LasR and RhlR) by their corresponding autoinducers (3-oxo-C12-homoserine lactone [HSL] and C4-HSL), which function as intercellular signals. We found that overexpression of relA activated the expression of rpoS in P. aeruginosa and led to premature, cell density-independent LasB elastase production. We therefore investigated the effects of the stringent response on quorum sensing. Both lasR and rhlR gene expression and autoinducer synthesis were prematurely activated during the stringent response induced by overexpression of relA. Premature expression of lasR and rhlR was also observed when relA was overexpressed in a PAO1 rpoSmutant. The stringent response induced by the amino acid analogue serine hydroxamate (SHX) also led to premature production of the 3-oxo-C12-HSL autoinducer. This response to SHX was absent in a PAO1 relA mutant. These findings suggest that the stringent response can activate the two quorum-sensing systems of P. aeruginosa independently of cell density.

scholarly journals Hierarchical Transcriptional Control of the LuxR Quorum-Sensing Regulon of Vibrio harveyi

2020 ◽  
Vol 202 (14) ◽  
Author(s):  
Ryan R. Chaparian ◽  
Alyssa S. Ball ◽  
Julia C. van Kessel
Keyword(s):  
Gene Expression ◽  
Transcription Factors ◽  
Quorum Sensing ◽  
Cell Density ◽  
Regulatory Networks ◽  
Vibrio Harveyi ◽  
Sugar Transport ◽  
Content Type ◽  
Second Tier

ABSTRACT In vibrios, quorum sensing controls hundreds of genes that are required for cell density-specific behaviors including bioluminescence, biofilm formation, competence, secretion, and swarming motility. The central transcription factor in the quorum-sensing pathway is LuxR/HapR, which directly regulates ∼100 genes in the >400-gene regulon of Vibrio harveyi. Among these directly controlled genes are 15 transcription factors, which we predicted would comprise the second tier in the hierarchy of the LuxR regulon. We confirmed that LuxR binds to the promoters of these genes in vitro and quantified the extent of LuxR activation or repression of transcript levels. Transcriptome sequencing (RNA-seq) indicates that most of these transcriptional regulators control only a few genes, with the exception of MetJ, which is a global regulator. The genes regulated by these transcription factors are predicted to be involved in methionine and thiamine biosynthesis, membrane stability, RNA processing, c-di-GMP degradation, sugar transport, and other cellular processes. These data support a hierarchical model in which LuxR directly regulates 15 transcription factors that drive the second level of the gene expression cascade to influence cell density-dependent metabolic states and behaviors in V. harveyi. IMPORTANCE Quorum sensing is important for survival of bacteria in nature and influences the actions of bacterial groups. In the relatively few studied examples of quorum-sensing-controlled genes, these genes are associated with competition or cooperation in complex microbial communities and/or virulence in a host. However, quorum sensing in vibrios controls the expression of hundreds of genes, and their functions are mostly unknown or uncharacterized. In this study, we identify the regulators of the second tier of gene expression in the quorum-sensing system of the aquaculture pathogen Vibrio harveyi. Our identification of regulatory networks and metabolic pathways controlled by quorum sensing can be extended and compared to other Vibrio species to understand the physiology, ecology, and pathogenesis of these organisms.

scholarly journals Characterization of a Legionella pneumophila relA Insertion Mutant and Roles of RelA and RpoS in Virulence Gene Expression

2002 ◽  
Vol 184 (1) ◽  
pp. 67-75 ◽  
Author(s):  
Tal Zusman ◽  
Ohad Gal-Mor ◽  
Gil Segal
Keyword(s):  
Gene Expression ◽  
Stationary Phase ◽  
Gene Product ◽  
Legionella Pneumophila ◽  
Type Strain ◽  
Wild Type Strain ◽  
Insertion Mutant ◽  
Wild Type ◽  
Intracellular Growth ◽  
Virulence Gene Expression

ABSTRACT To investigate the involvement of RelA in the regulation of Legionella pneumophila virulence, a deletion substitution was constructed in the relA gene. The relA knockout resulted in an undetectable level of ppGpp in the cells during the stationary phase, but the original level was restored when the relA gene product was supplied on a plasmid. The effect of the relA mutation was examined with two systems that are known to be expressed during the stationary phase in L. pneumophila. Pigment production was found to be dependent on the relA gene product, and only one-half as much pigment was produced by the relA mutant as by the wild-type strain. Flagellum gene expression was also found to be dependent on the relA gene product, as determined with a flaA::lacZ fusion. However, the relA gene product was found to be dispensable for intracellular growth both in HL-60-derived human macrophages and in the protozoan host Acanthamoeba castellanii. To determine the involvement of the relA gene product in expression of L. pneumophila genes required for intracellular growth (icm/dot genes), nine icm::lacZ fusions were constructed, and expression of these fusions in the wild-type strain was compared with their expression in relA mutant strains. Expression of only one of the icm::lacZ fusions was moderately reduced in the relA mutant strain. Expression of the nine icm::lacZ fusions was also examined in a strain containing an insertion in the gene that codes for the stationary-phase sigma factor RpoS, and similar results were obtained. We concluded that RelA is dispensable for intracellular growth of L. pneumophila in the two hosts examined and that both RelA and RpoS play minor roles in L. pneumophila icm/dot gene expression.

scholarly journals In VivoProgrammed Gene Expression Based on Artificial Quorum Networks

2015 ◽  
Vol 81 (15) ◽  
pp. 4984-4992 ◽  
Author(s):  
Teng Chu ◽  
Yajun Huang ◽  
Mingyu Hou ◽  
Qiyao Wang ◽  
Jingfan Xiao ◽  
...  
Keyword(s):  
Gene Expression ◽  
Quorum Sensing ◽  
Cell Density ◽  
Protective Antigen ◽  
Expression System ◽  
Edwardsiella Tarda ◽  
Content Type ◽  
Genetic Components ◽  
Wide Range

ABSTRACTThe quorum sensing (QS) system, as a well-functioning population-dependent gene switch, has been widely applied in many gene circuits in synthetic biology. In our work, an efficient cell density-controlled expression system (QS) was established via engineering of theVibrio fischeri luxI-luxRquorum sensing system. In order to achievein vivoprogrammed gene expression, a synthetic binary regulation circuit (araQS) was constructed by assembling multiple genetic components, including the quorum quenching protein AiiA and the arabinose promoter ParaBAD, into the QS system.In vitroexpression assays verified that the araQS system was initiated only in the absence of arabinose in the medium at a high cell density.In vivoexpression assays confirmed that the araQS system presented anin vivo-triggered and cell density-dependent expression pattern. Furthermore, the araQS system was demonstrated to function well in different bacteria, indicating a wide range of bacterial hosts for use. To explore its potential applicationsin vivo, the araQS system was used to control the production of a heterologous protective antigen in an attenuatedEdwardsiella tardastrain, which successfully evoked efficient immune protection in a fish model. This work suggested that the araQS system could program bacterial expressionin vivoand might have potential uses, including, but not limited to, bacterial vector vaccines.

scholarly journals LuxT is a Global Regulator of Low-Cell Density Behaviors Including Type III Secretion, Siderophore Production, and Aerolysin Secretion in Vibrio harveyi

2021 ◽  
Author(s):  
Michaela J. Eickhoff ◽  
Chenyi Fei ◽  
Jian-Ping Cong ◽  
Bonnie L. Bassler
Keyword(s):  
Gene Expression ◽  
Transcription Factor ◽  
Cell Density ◽  
Type Iii Secretion ◽  
Siderophore Production ◽  
Communication Process ◽  
Global Regulator ◽  
Type Iii ◽  
Regulatory Cascade ◽  
Low Cell Density

Quorum sensing (QS) is a chemical communication process in which bacteria produce, release, and detect extracellular signaling molecules called autoinducers. Via combined transcriptional and post-transcriptional regulatory mechanisms, QS allows bacteria to collectively alter gene expression on a population-wide scale. Recently, the LuxT transcription factor was shown to control V. harveyiqrr1, encoding the Qrr1 small RNA that functions at the core of the QS regulatory cascade. Here, we use RNA-Sequencing to reveal that, beyond control of qrr1, LuxT is a global regulator of 414 V. harveyi genes including those involved in type III secretion, siderophore production, and aerolysin toxin biosynthesis. Importantly, LuxT directly represses swrZ, encoding a transcription factor, and LuxT control of type III secretion, siderophore, and aerolysin genes occurs by two mechanisms, one that is SwrZ-dependent and one that is SwrZ-independent. All of these target genes specify QS-controlled behaviors that are enacted when V. harveyi is at low cell density. Thus, LuxT and SwrZ function in parallel with QS to drive particular low cell density behaviors. Phylogenetic analyses reveal that luxT is highly conserved among Vibrionaceae, but swrZ is less well conserved. In a test case to examine the relationship between LuxT and SwrZ, we find that in Aliivibrio fischeri, LuxT also functions as a swrZ repressor, and LuxT activates A. fischeri siderophore production via swrZ repression. Our results indicate that LuxT is a major regulator among Vibrionaceae, and, in the species that also possess swrZ, LuxT functions with SwrZ to control gene expression.

scholarly journals Global H-NS counter-silencing by LuxR activates quorum sensing gene expression

2019 ◽  
Author(s):  
Ryan R Chaparian ◽  
Minh L N Tran ◽  
Laura C Miller Conrad ◽  
Douglas B Rusch ◽  
Julia C van Kessel
Keyword(s):  
Gene Expression ◽  
Quorum Sensing ◽  
Cell Communication ◽  
Binding Activity ◽  
Primary Role ◽  
Dna Binding Activity ◽  
Cellular Behaviors ◽  
A Cell ◽  
Promoter Dna

Abstract Bacteria coordinate cellular behaviors using a cell–cell communication system termed quorum sensing. In Vibrio harveyi, the master quorum sensing transcription factor LuxR directly regulates >100 genes in response to changes in population density. Here, we show that LuxR derepresses quorum sensing loci by competing with H-NS, a global transcriptional repressor that oligomerizes on DNA to form filaments and bridges. We first identified H-NS as a repressor of bioluminescence gene expression, for which LuxR is a required activator. In an hns deletion strain, LuxR is no longer necessary for transcription activation of the bioluminescence genes, suggesting that the primary role of LuxR is to displace H-NS to derepress gene expression. Using RNA-seq and ChIP-seq, we determined that H-NS and LuxR co-regulate and co-occupy 28 promoters driving expression of 63 genes across the genome. ChIP-PCR assays show that as autoinducer concentration increases, LuxR protein accumulates at co-occupied promoters while H-NS protein disperses. LuxR is sufficient to evict H-NS from promoter DNA in vitro, which is dependent on LuxR DNA binding activity. From these findings, we propose a model in which LuxR serves as a counter-silencer at H-NS-repressed quorum sensing loci by disrupting H-NS nucleoprotein complexes that block transcription.

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