scholarly journals The Cpx Envelope Stress Response Is Controlled by Amplification and Feedback Inhibition

1999 ◽  
Vol 181 (17) ◽  
pp. 5263-5272 ◽  
Author(s):  
Tracy L. Raivio ◽  
Daniel L. Popkin ◽  
Thomas J. Silhavy
Keyword(s):  
Escherichia Coli ◽  
Stress Response ◽  
Virulence Factors ◽  
Histidine Kinase ◽  
Feedback Inhibition ◽  
Response Regulator ◽  
Regulatory System ◽  
Concomitant Increase ◽  
Envelope Stress ◽  
Two Component

ABSTRACT In Escherichia coli, the Cpx two-component regulatory system activates expression of protein folding and degrading factors in response to misfolded proteins in the bacterial envelope (inner membrane, periplasm, and outer membrane). It is comprised of the histidine kinase CpxA and the response regulator CpxR. This response plays a role in protection from stresses, such as elevated pH, as well as in the biogenesis of virulence factors. Here, we show that the Cpx periplasmic stress response is subject to amplification and repression through positive and negative autofeedback mechanisms. Western blot and operon fusion analyses demonstrated that the cpxRA operon is autoactivated. Conditions that lead to elevated levels of phosphorylated CpxR cause a concomitant increase in transcription ofcpxRA. Conversely, overproduction of CpxP, a small, Cpx-regulated protein of previously unknown function, represses the regulon and can block activation of the pathway. This repression is dependent on an intact CpxA sensing domain. The ability to autoactivate and then subsequently repress allows for a temporary amplification of the Cpx response that may be important in rescuing cells from transitory stresses and cueing the appropriately timed elaboration of virulence factors.

scholarly journals Fumarate Regulation of Gene Expression in Escherichia coli by the DcuSR (dcuSR Genes) Two-Component Regulatory System

1998 ◽  
Vol 180 (20) ◽  
pp. 5421-5425 ◽  
Author(s):  
Evelyn Zientz ◽  
Johannes Bongaerts ◽  
Gottfried Unden
Keyword(s):  
Escherichia Coli ◽  
Histidine Kinase ◽  
Response Regulator ◽  
Regulation Of Gene Expression ◽  
Regulatory System ◽  
E Coli ◽  
Expression In Escherichia Coli ◽  
Two Component ◽  
Genes Encoding ◽  
Periplasmic Domain

ABSTRACT In Escherichia coli the genes encoding the anaerobic fumarate respiratory system are transcriptionally regulated by C4-dicarboxylates. The regulation is effected by a two-component regulatory system, DcuSR, consisting of a sensory histidine kinase (DcuS) and a response regulator (DcuR). DcuS and DcuR are encoded by the dcuSR genes (previouslyyjdHG) at 93.7 min on the calculated E. coli map. Inactivation of the dcuR anddcuS genes caused the loss of C4-dicarboxylate-stimulated synthesis of fumarate reductase (frdABCD genes) and of the anaerobic fumarate-succinate antiporter DcuB (dcuB gene). DcuS is predicted to contain a large periplasmic domain as the supposed site for C4-dicarboxylate sensing. Regulation by DcuR and DcuS responded to the presence of the C4-dicarboxylates fumarate, succinate, malate, aspartate, tartrate, and maleate. Since maleate is not taken up by the bacteria under these conditions, the carboxylates presumably act from without. Genes of the aerobic C4-dicarboxylate pathway encoding succinate dehydrogenase (sdhCDAB) and the aerobic succinate carrier (dctA) are only marginally or negatively regulated by the DcuSR system. The CitAB two-component regulatory system, which is highly similar to DcuSR, had no effect on C4-dicarboxylate regulation of any of the genes.

scholarly journals Characterization of the CpxRA Regulon in Haemophilus ducreyi

2010 ◽  
Vol 78 (11) ◽  
pp. 4779-4791 ◽  
Author(s):  
Maria Labandeira-Rey ◽  
Chad A. Brautigam ◽  
Eric J. Hansen
Keyword(s):  
Stress Response ◽  
Response Regulator ◽  
Cell Envelope ◽  
Bacterial Species ◽  
Regulatory System ◽  
Promoter Regions ◽  
Mobility Shift ◽  
Envelope Stress ◽  
Two Component ◽  
Genes Encoding

ABSTRACT The H aemophilus ducreyi 35000HP genome encodes a homolog of the CpxRA two-component cell envelope stress response system originally characterized in E scherichia coli. CpxR, the cytoplasmic response regulator, was shown previously to be involved in repression of the expression of the lspB-lspA2 operon (M. Labandeira-Rey, J. R. Mock, and E. J. Hansen, Infect. Immun. 77:3402-3411, 2009). In the present study, the H. ducreyi CpxR and CpxA proteins were shown to closely resemble those of other well-studied bacterial species. A cpxA deletion mutant and a CpxR-overexpressing strain were used to explore the extent of the CpxRA regulon. DNA microarray and real-time reverse transcriptase (RT) PCR analyses indicated several potential regulatory targets for the H. ducreyi CpxRA two-component regulatory system. Electrophoretic mobility shift assays (EMSAs) were used to prove that H. ducreyi CpxR interacted with the promoter regions of genes encoding both known and putative virulence factors of H. ducreyi, including the lspB-lspA2 operon, the flp operon, and dsrA. Interestingly, the use of EMSAs also indicated that H. ducreyi CpxR did not bind to the promoter regions of several genes predicted to encode factors involved in the cell envelope stress response. Taken together, these data suggest that the CpxRA system in H. ducreyi, in contrast to that in E. coli, may be involved primarily in controlling expression of genes not involved in the cell envelope stress response.

scholarly journals The Escherichia coli CpxA-CpxR Envelope Stress Response System Regulates Expression of the Porins OmpF and OmpC

2005 ◽  
Vol 187 (16) ◽  
pp. 5723-5731 ◽  
Author(s):  
Eric Batchelor ◽  
Don Walthers ◽  
Linda J. Kenney ◽  
Mark Goulian
Keyword(s):  
Escherichia Coli ◽  
Stress Response ◽  
Response Regulator ◽  
Gene Encoding ◽  
Fluorescent Reporter ◽  
Response System ◽  
Dnase I Footprinting ◽  
Envelope Stress ◽  
Two Component ◽  
Stress Response System

ABSTRACT We performed transposon mutagenesis of a two-color fluorescent reporter strain to identify new regulators of the porin genes ompF and ompC in Escherichia coli. Screening of colonies by fluorescence microscopy revealed numerous mutants that exhibited interesting patterns of porin expression. One mutant harbored an insertion in the gene encoding the histidine kinase CpxA, the sensor for a two-component signaling system that responds to envelope stress. The cpxA mutant exhibited increased transcription of ompC and a very strong decrease in transcription of ompF under conditions in which acetyl phosphate levels were high. Subsequent genetic analysis revealed that this phenotype is dependent on phosphorylation of the response regulator CpxR and that activation of CpxA in wild-type cells results in similar regulation of porin expression. Using DNase I footprinting, we demonstrated that CpxR binds upstream of both the ompF and ompC promoters. It thus appears that two distinct two-component systems, CpxA-CpxR and EnvZ-OmpR, converge at the porin promoters. Within the context of envelope stress, outer membrane beta-barrel proteins have generally been associated with the sigma E pathway. However, at least for the classical porins OmpF and OmpC, our results show that the Cpx envelope stress response system plays a role in regulating their expression.

scholarly journals Localization and Cellular Amounts of the WalRKJ (VicRKX) Two-Component Regulatory System Proteins in Serotype 2 Streptococcus pneumoniae

2010 ◽  
Vol 192 (17) ◽  
pp. 4388-4394 ◽  
Author(s):  
Kyle J. Wayne ◽  
Lok-To Sham ◽  
Ho-Ching T. Tsui ◽  
Alina D. Gutu ◽  
Skye M. Barendt ◽  
...  
Keyword(s):  
Streptococcus Pneumoniae ◽  
Histidine Kinase ◽  
Cellular Localization ◽  
Immunofluorescence Microscopy ◽  
Response Regulator ◽  
Regulatory System ◽  
Respiratory Pathogen ◽  
Cell Periphery ◽  
Gram Positive Bacteria ◽  
Two Component

ABSTRACT The WalRK two-component regulatory system coordinates gene expression that maintains cell wall homeostasis and responds to antibiotic stress in low-GC Gram-positive bacteria. Phosphorylated WalR (VicR) of the major human respiratory pathogen Streptococcus pneumoniae (WalR Spn ) positively regulates transcription of several surface virulence genes and, most critically, pcsB, which encodes an essential cell division protein. Despite numerous studies of several species, little is known about the signals sensed by the WalK histidine kinase or the function of the WalJ ancillary protein encoded in the walRKSpn operon. To better understand the functions of the WalRKJ Spn proteins in S. pneumoniae, we performed experiments to determine their cellular localization and amounts. In contrast to WalK from Bacillus subtilis (WalK Bsu ), which is localized at division septa, immunofluorescence microscopy showed that WalK Spn is distributed throughout the cell periphery. WalJ Spn is also localized to the cell surface periphery, whereas WalR Spn was found to be localized in the cytoplasm around the nucleoid. In fractionation experiments, WalR Spn was recovered from the cytoplasmic fraction, while WalK Spn and the majority of WalJ Spn were recovered from the cell membrane fraction. This fractionation is consistent with the localization patterns observed. Lastly, we determined the cellular amounts of WalRKJ Spn by quantitative Western blotting. The WalR Spn response regulator is relatively abundant and present at levels of ≈6,200 monomers per cell, which are ≈14-fold greater than the amount of the WalK Spn histidine kinase, which is present at ≈460 dimers (920 monomers) per cell. We detected ≈1,200 monomers per cell of WalJ Spn ancillary protein, similar to the amount of WalK Spn .

scholarly journals Signal Detection and Target Gene Induction by the CpxRA Two-Component System

2003 ◽  
Vol 185 (8) ◽  
pp. 2432-2440 ◽  
Author(s):  
Patricia A. DiGiuseppe ◽  
Thomas J. Silhavy
Keyword(s):  
Signal Transduction ◽  
Feedback Inhibition ◽  
Response Regulator ◽  
Negative Regulator ◽  
Outer Membrane Lipoprotein ◽  
Envelope Stress ◽  
Two Component ◽  
Two Component Signal Transduction ◽  
Cpx Pathway ◽  
Target Gene Induction

ABSTRACT The Cpx pathway is a two-component signal transduction system that senses a variety of envelope stresses, including misfolded proteins, and responds by upregulating periplasmic folding and trafficking factors. CpxA resides in the inner membrane and has both kinase and phosphatase activities. CpxR, the response regulator, mediates a response by activating transcription of stress-combative genes. Signal transduction is subject to feedback inhibition via regulon member CpxP and autoamplification. Recently, it was shown that the Cpx pathway is also upregulated when cells adhere to hydrophobic surfaces and that this response is dependent on the outer membrane lipoprotein NlpE. Here we show that while NlpE is required for induction of the Cpx pathway by adhesion, induction by envelope stress and during growth is NlpE independent. We show that while all of the envelope stresses tested induce the Cpx pathway in a manner that is dependent on the periplasmic domain of CpxA, induction during growth is independent of CpxA. Therefore, we propose that the Cpx pathway can sense inducing cues that enter the signaling pathway at three distinct points. Although CpxP is not required for induction of the Cpx pathway, we show that its activity as a negative regulator of CpxA is inactivated by envelope stress. Moreover, the cpxP promoter is more inducible than any other regulon member tested. Consistent with these results, we suggest that CpxP performs a second function, most likely that of a chaperone. Finally, we show that two Cpx-regulated genes are differentially upregulated in response to different envelope stresses, suggesting the existence of three stress-responsive systems.

scholarly journals Succinoglycan Production by Rhizobium meliloti Is Regulated through the ExoS-ChvI Two-Component Regulatory System

1998 ◽  
Vol 180 (1) ◽  
pp. 20-26 ◽  
Author(s):  
Hai-Ping Cheng ◽  
Graham C. Walker
Keyword(s):  
Histidine Kinase ◽  
Cytoplasmic Membrane ◽  
Rhizobium Meliloti ◽  
Response Regulator ◽  
Regulatory System ◽  
Kinase Domain ◽  
Cloning And Sequencing ◽  
System A ◽  
Two Component ◽  
Close Homolog

ABSTRACT The Rhizobium meliloti exoS gene is involved in regulating the production of succinoglycan, which plays a crucial role in the establishment of the symbiosis between R. melilotiRm1021 and its host plant, alfalfa. TheexoS96::Tn5 mutation causes the upregulation of the succinoglycan biosynthetic genes, thereby resulting in the overproduction of succinoglycan. Through cloning and sequencing, we found that the exoS gene is a close homolog of theAgrobacterium tumefaciens chvG gene, which has been proposed to encode the sensor protein of the ChvG-ChvI two-component regulatory system, a member of the EnvZ-OmpR family. Further analyses revealed the existence of a newly discovered A. tumefaciens chvI homolog located just upstream of the R. meliloti exoS gene. R. meliloti ChvI may serve as the response regulator of ExoS in a two-component regulatory system. By using ExoS-specific antibodies, it was found that the ExoS protein cofractionated with membrane proteins, suggesting that it is located in the cytoplasmic membrane. By using the same antibodies, it was shown that the exoS96::Tn5 allele encodes an N-terminal truncated derivative of ExoS. The cytoplasmic histidine kinase domain of ExoS was expressed in Escherichia coli and purified, as was the R. meliloti ChvI protein. The ChvI protein autophosphorylated in the presence of acetylphosphate, and the ExoS cytoplasmic domain fragment autophosphorylated at a histidine residue in the presence of ATP. The ChvI protein was phosphorylated in the presence of ATP only when the histidine kinase domain of ExoS was also present. We propose a model for regulation of succinoglycan production by R. meliloti through the ExoS-ChvI two-component regulatory system.

scholarly journals The BaeSR Two-Component Regulatory System Mediates Resistance to Condensed Tannins in Escherichia coli

2007 ◽  
Vol 74 (2) ◽  
pp. 535-539 ◽  
Author(s):  
Erwin G. Zoetendal ◽  
Alexandra H. Smith ◽  
Monica A. Sundset ◽  
Roderick I. Mackie
Keyword(s):  
Escherichia Coli ◽  
Cell Envelope ◽  
Expression Profiles ◽  
Stress Protein ◽  
Gene Expression Profiles ◽  
Regulatory System ◽  
Multidrug Transporter ◽  
Resistance Strategies ◽  
Envelope Stress ◽  
Two Component

ABSTRACT The gene expression profiles of Escherichia coli strains grown anaerobically with or without Acacia mearnsii (black wattle) extract were compared to identify tannin resistance strategies. The cell envelope stress protein gene spy and the multidrug transporter-encoding operon mdtABCD, both under the control of the BaeSR two-component regulatory system, were significantly up-regulated in the presence of tannins. BaeSR mutants were more tannin sensitive than their wild-type counterparts.

scholarly journals Mutations Affecting HVO_1357 or HVO_2248 Cause Hypermotility in Haloferax volcanii, Suggesting Roles in Motility Regulation

Genes ◽  
2020 ◽  
Vol 12 (1) ◽  
pp. 58
Author(s):  
Michiyah Collins ◽  
Simisola Afolayan ◽  
Aime B. Igiraneza ◽  
Heather Schiller ◽  
Elise Krespan ◽  
...  
Keyword(s):  
Histidine Kinase ◽  
Response Regulator ◽  
Regulatory System ◽  
Haloferax Volcanii ◽  
Adjacent Gene ◽  
Transposon Insertion ◽  
Regulatory Systems ◽  
Two Component ◽  
Motility Regulation ◽  
Transposon Insertions

Motility regulation plays a key role in prokaryotic responses to environmental stimuli. Here, we used a motility screen and selection to isolate hypermotile Haloferax volcanii mutants from a transposon insertion library. Whole genome sequencing revealed that hypermotile mutants were predominantly affected in two genes that encode HVO_1357 and HVO_2248. Alterations of these genes comprised not only transposon insertions but also secondary genome alterations. HVO_1357 contains a domain that was previously identified in the regulation of bacteriorhodopsin transcription, as well as other domains frequently found in two-component regulatory systems. The genes adjacent to hvo_1357 encode a sensor box histidine kinase and a response regulator, key players of a two-component regulatory system. None of the homologues of HVO_2248 have been characterized, nor does it contain any of the assigned InterPro domains. However, in a significant number of Haloferax species, the adjacent gene codes for a chemotaxis receptor/transducer. Our results provide a foundation for characterizing the root causes underlying Hfx. volcanii hypermotility.

scholarly journals The BaeSR Two-Component Regulatory System Activates Transcription of the yegMNOB (mdtABCD) Transporter Gene Cluster in Escherichia coli and Increases Its Resistance to Novobiocin and Deoxycholate

2002 ◽  
Vol 184 (15) ◽  
pp. 4168-4176 ◽  
Author(s):  
Natalya Baranova ◽  
Hiroshi Nikaido
Keyword(s):  
Escherichia Coli ◽  
Response Regulator ◽  
Regulatory System ◽  
Major Facilitator Superfamily ◽  
Multicopy Plasmid ◽  
Active Efflux ◽  
Membrane Fusion Protein ◽  
Two Component ◽  
Major Facilitator

ABSTRACT Screening of random fragments of Escherichia coli genomic DNA for their ability to increase the novobiocin resistance of a hypersusceptible ΔacrAB mutant resulted in the isolation of a plasmid containing baeR, which codes for the response regulator of the two-component regulatory system BaeSR. When induced for expression, baeR cloned in multicopy plasmid pTrc99A significantly increased the resistance of the ΔacrAB host strain to novobiocin (16-fold) and to deoxycholate (8-fold). Incubation of cells with novobiocin followed by a chromatographic assay for intracellular drug showed that overproduced BaeR decreased drastically the drug accumulation, presumably via increased active efflux. The genes baeSR are part of a putative operon, yegMNOB baeSR. Direct binding of BaeR to the yegM promoter was demonstrated in vitro by gel retardation assay. The gene yegB, which codes for a major facilitator superfamily transporter, was not necessary for increased resistance, but deletion of yegO or an in-frame deletion of yegN, both of which code for resistance-nodulation-cell division-type multidrug transporters, abolished the BaeR-induced increase in resistance. It is likely that both YegN and YegO produce a complex(es) with the membrane fusion protein family member YegM and pump out novobiocin and deoxycholate. We accordingly propose to rename yegMNOB as mdtABCD (mdt for multidrug transporter). Finally, the expression of two other genes, yicO and ygcL, was shown to be regulated by BaeR, but it is not known if they play any roles in resistance.

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