Global chromosome topology and the two-component systems in concerted manner regulate transcription in Streptomyces

2021 ◽  
Author(s):  
Martyna Gongerowska-Jac ◽  
Marcin Jan Szafran ◽  
Jakub Mikołajczyk ◽  
Justyna Szymczak ◽  
Magdalena Bartyńska ◽  
...  
Keyword(s):  
Topoisomerase I ◽  
Dna Supercoiling ◽  
Complex Life Cycle ◽  
Two Component System ◽  
Bacterial Gene ◽  
Component System ◽  
Global Regulators ◽  
Component Systems ◽  
Two Component ◽  
Two Component Systems

Bacterial gene expression is controlled at multiple levels, with chromosome supercoiling being one of the most global regulators. Global DNA supercoiling is maintained by the orchestrated action of topoisomerases. In Streptomyces, mycelial soil bacteria with a complex life cycle, topoisomerase I depletion led to elevated chromosome supercoiling, changed expression of significant fraction of genes, delayed growth and blocked sporulation. To identify supercoiling-induced sporulation regulators, we searched for S. coelicolor transposon mutants that were able to restore sporulation despite high chromosome supercoiling. We established that transposon insertion in genes encoding a novel two-component system named SatKR reversed the sporulation blockage resulting from topoisomerase I depletion. Transposition in satKR abolished the transcriptional induction of the genes within the so-called supercoiling-hypersensitive cluster (SHC). Moreover, we found that activated SatR also induced the same set of SHC genes under normal supercoiling conditions. We determined that the expression of genes in this region impacted S. coelicolor growth and sporulation. Interestingly, among the associated products is another two-component system (SitKR), indicating the potential for cascading regulatory effects driven by the SatKR and SitKR two-component systems. Thus, we demonstrated the concerted activity of chromosome supercoiling and a hierarchical two-component signalling system that impacts gene activity governing Streptomyces growth and sporulation.

Monte Carlo Simulation of Electronic Excitation Migrationand Trappingin Disordered Two-Component Systems. A Comparison with Analytical Theories

1989 ◽  
Vol 44 (4) ◽  
pp. 257-261 ◽  
Author(s):  
Sławomir Błonski ◽  
Czesław Bojarski
Keyword(s):  
Monte Carlo ◽  
Electronic Excitation ◽  
Two Component System ◽  
Component System ◽  
Steady State Fluorescence ◽  
Donor And Acceptor ◽  
Component Systems ◽  
Two Component ◽  
Two Component Systems ◽  
Viscous Solution

Abstract Monte Carlo simulations of quantum yield and anisotropy of fluorescence in two-component systems have been conducted with various donor and acceptor concentrations and Förster radii ratios RDAO/RDDO. The influence of excitation migration and trapping on the fluorescence of the viscous solution has been considered. The results of the simulations have shown that steady-state fluorescence of a two-component system depends on the RDAO/RDDO ratio as predicted in LAF theory.

scholarly journals The two-component system CopRS maintains femtomolar levels of free copper in the periplasm of Pseudomonas aeruginosa using a phosphatase-based mechanism

2020 ◽  
Author(s):  
Lorena Novoa-Aponte ◽  
Fernando C. Soncini ◽  
José M. Argüello
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Phosphatase Activity ◽  
Response Regulator ◽  
Redox Enzymes ◽  
Two Component System ◽  
Component System ◽  
Component Systems ◽  
Two Component ◽  
Two Component Systems ◽  
Systems Control

ABSTRACTTwo component systems control periplasmic Cu+ homeostasis in Gram-negative bacteria. In characterized systems such as Escherichia coli CusRS, upon Cu+ binding to the periplasmic sensing domain of CusS, a cytoplasmic phosphotransfer domain phosphorylates the response regulator CusR. This drives the expression of efflux transporters, chaperones, and redox enzymes to ameliorate metal toxic effects. Here, we show that the Pseudomonas aeruginosa two component sensor histidine kinase CopS exhibits a Cu-dependent phosphatase activity that maintains a non-phosphorylated CopR when the periplasmic Cu levels are below its activation threshold. Upon Cu+ binding to the sensor, the phosphatase activity is blocked and the phosphorylated CopR activates transcription of the CopRS regulon. Supporting the model, mutagenesis experiments revealed that the ΔcopS strain showed constitutive high expression of the CopRS regulon, lower intracellular Cu+ levels, and larger Cu tolerance when compared to wild type cells. The invariant phospho-acceptor residue His235 of CopS was not required for the phosphatase activity itself, but necessary for its Cu-dependency. To sense the metal, the periplasmic domain of CopS binds two Cu+ ions at its dimeric interface. Homology modeling of CopS based on CusS structure (four Ag+ binding sites) clearly explains the different binding stoichiometries in both systems. Interestingly, CopS binds Cu+/2+ with 30 × 10−15 M affinities, pointing to the absence of free (hydrated) Cu+/2+ in the periplasm.IMPORTANCECopper is a micronutrient required as cofactor in redox enzymes. When free, copper is toxic, mismetallating proteins, and generating damaging free radicals. Consequently, copper overload is a strategy that eukaryotic cells use to combat pathogens. Bacteria have developed copper sensing transcription factors to control copper homeostasis. The cell envelope is the first compartment that has to cope with copper stress. Dedicated two component systems control the periplasmic response to metal overload. This manuscript shows that the copper sensing two component system present in Pseudomonadales exhibits a signal-dependent phosphatase activity controlling the activation of the response regulator, distinct from previously described periplasmic Cu sensors. Importantly, the data show that the sensor is activated by copper levels compatible with the absence of free copper in the cell periplasm. This emphasizes the diversity of molecular mechanisms that have evolved in various bacteria to manage the copper cellular distribution.

scholarly journals The ChrA-ChrS and HrrA-HrrS Signal Transduction Systems Are Required for Activation of the hmuO Promoter and Repression of the hemA Promoter in Corynebacterium diphtheriae

2007 ◽  
Vol 75 (5) ◽  
pp. 2421-2431 ◽  
Author(s):  
Lori A. Bibb ◽  
Carey A. Kunkle ◽  
Michael P. Schmitt
Keyword(s):  
Corynebacterium Diphtheriae ◽  
Heme Iron ◽  
Dependent Manner ◽  
Two Component System ◽  
Component System ◽  
Regulatory Systems ◽  
Component Systems ◽  
Two Component ◽  
Genes Encoding ◽  
Two Component Systems

ABSTRACT Transcription of the Corynebacterium diphtheriae hmuO gene, which encodes a heme oxygenase involved in heme iron utilization, is activated in a heme- or hemoglobin-dependent manner in part by the two-component system ChrA-ChrS. Mutation of either the chrA or the chrS gene resulted in a marked reduction of hemoglobin-dependent activation at the hmuO promoter in C. diphtheriae; however, it was observed that significant levels of hemoglobin-dependent expression were maintained in the mutants, suggesting that an additional activator is involved in regulation. A BLAST search of the C. diphtheriae genome sequence revealed a second two-component system, encoded by DIP2268 and DIP2267, that shares similarity with ChrS and ChrA, respectively; we have designated these genes hrrS (DIP2268) and hrrA (DIP2267). Analysis of hmuO promoter expression demonstrated that hemoglobin-dependent activity was fully abolished in strains from which both the chrA-chrS and the hrrA-hrrS two-component systems were deleted. Similarly, deletion of the sensor kinase genes chrS and hrrS or the genes encoding both of the response regulators chrA and hrrA also eliminated hemoglobin-dependent activation at the hmuO promoter. We also show that the regulators ChrA-ChrS and HrrA-HrrS are involved in the hemoglobin-dependent repression of the promoter upstream of hemA, which encodes a heme biosynthesis enzyme. Evidence for cross talk between the ChrA-ChrS and HrrA-HrrS systems is presented. In conclusion, these findings demonstrate that the ChrA-ChrS and HrrA-HrrS regulatory systems are critical for full hemoglobin-dependent activation at the hmuO promoter and also suggest that these two-component systems are involved in the complex mechanism of the regulation of heme homeostasis in C. diphtheriae.

scholarly journals Genome-Wide Analysis of Two-Component Systems and Prediction of Stress-Responsive Two-Component System Members in Soybean

DNA Research ◽  
2010 ◽  
Vol 17 (5) ◽  
pp. 303-324 ◽  
Author(s):  
K. Mochida ◽  
T. Yoshida ◽  
T. Sakurai ◽  
K. Yamaguchi-Shinozaki ◽  
K. Shinozaki ◽  
...  
Keyword(s):  
Two Component System ◽  
Component System ◽  
Genome Wide Analysis ◽  
Genome Wide ◽  
Component Systems ◽  
Two Component ◽  
Two Component Systems

Phase transitions in two-component systems

1959 ◽  
Vol 249 (1258) ◽  
pp. 335-345
Keyword(s):  
Phase Transitions ◽  
Binary Mixtures ◽  
Fourier Transforms ◽  
Two Component System ◽  
Component System ◽  
System A ◽  
Component Systems ◽  
Two Component ◽  
Two Component Systems

The Born-Green equations have been generalized to binary mixtures, and solutions in terms of Fourier transforms have been found. The singularities of the solution are believed to correspond to the point at which condensation first occurs in the two component system. A plot of this temperature against density for three mole fractions is shown.

scholarly journals The Legionella pneumophila LetA/LetS Two-Component System Exhibits Rheostat-Like Behavior

2010 ◽  
Vol 78 (6) ◽  
pp. 2571-2583 ◽  
Author(s):  
Rachel L. Edwards ◽  
Matthieu Jules ◽  
Tobias Sahr ◽  
Carmen Buchrieser ◽  
Michele S. Swanson
Keyword(s):  
Gene Expression ◽  
Legionella Pneumophila ◽  
Profile Analysis ◽  
Metabolic Stress ◽  
Two Component System ◽  
Component System ◽  
Component Systems ◽  
Two Component ◽  
Flagellar Regulon ◽  
Two Component Systems

ABSTRACT When confronted with metabolic stress, replicative Legionella pneumophila bacteria convert to resilient, infectious cells equipped for transmission. Differentiation is promoted by the LetA/LetS two-component system, which belongs to a family of signal-transducing proteins that employ a four-step phosphorelay to regulate gene expression. Histidine 307 of LetS was essential to switch on the transmission profile, but a threonine substitution at position 311 (T311M) suggested a rheostat-like function. The letS(T311M) bacteria resembled the wild type (WT) for some traits and letS null mutants for others, whereas they displayed intermediate levels of infectivity, cytotoxicity, and lysosome evasion. Although only 30 to 50% of letS(T311M) mutants became motile, flow cytometry determined that every cell eventually activated the flagellin promoter to WT levels, but expression was delayed. Likewise, letS(T311M) mutants exhibited delayed induction of RsmY and RsmZ, regulatory RNAs that relieve CsrA repression of transmission traits. Transcriptional profile analysis revealed that letS(T311M) mutants expressed the flagellar regulon and multiple other transmissive-phase loci at a higher cell density than the WT. Accordingly, we postulate that the letS(T311M) mutant may relay phosphate less efficiently than the WT LetS sensor protein, leading to sluggish gene expression and a variety of phenotypic profiles. Thus, as first described for BvgA/BvgS, rather than acting as on/off switches, this family of two-component systems exhibit rheostat activity that likely confers versatility as microbes adapt to fluctuating environments.

scholarly journals The Two-Component System CopRS Maintains Subfemtomolar Levels of Free Copper in the Periplasm of Pseudomonas aeruginosa Using a Phosphatase-Based Mechanism

mSphere ◽  
2020 ◽  
Vol 5 (6) ◽  
pp. e01193-20
Author(s):  
Lorena Novoa-Aponte ◽  
Cheng Xu ◽  
Fernando C. Soncini ◽  
José M. Argüello
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Phosphatase Activity ◽  
Response Regulator ◽  
Redox Enzymes ◽  
Two Component System ◽  
Component System ◽  
Content Type ◽  
Component Systems ◽  
Two Component ◽  
Two Component Systems

ABSTRACTTwo-component systems control periplasmic Cu+ homeostasis in Gram-negative bacteria. In characterized systems such as Escherichia coli CusRS, upon Cu+ binding to the periplasmic sensing region of CusS, a cytoplasmic phosphotransfer domain of the sensor phosphorylates the response regulator CusR. This drives the expression of efflux transporters, chaperones, and redox enzymes to ameliorate metal toxic effects. Here, we show that the Pseudomonas aeruginosa two-component sensor histidine kinase CopS exhibits a Cu-dependent phosphatase activity that maintains CopR in a nonphosphorylated state when the periplasmic Cu levels are below the activation threshold of CopS. Upon Cu+ binding to the sensor, the phosphatase activity is blocked and the phosphorylated CopR activates transcription of the CopRS regulon. Supporting the model, mutagenesis experiments revealed that the ΔcopS strain exhibits maximal expression of the CopRS regulon, lower intracellular Cu+ levels, and increased Cu tolerance compared to wild-type cells. The invariant phosphoacceptor residue His235 of CopS was not required for the phosphatase activity itself but was necessary for its Cu dependency. To sense the metal, the periplasmic domain of CopS binds two Cu+ ions at its dimeric interface. Homology modeling of CopS based on CusS structure (four Ag+ binding sites) clearly supports the different binding stoichiometries in the two systems. Interestingly, CopS binds Cu+/2+ with 3 × 10−14 M affinity, pointing to the absence of free (hydrated) Cu+/2+ in the periplasm.IMPORTANCE Copper is a micronutrient required as cofactor in redox enzymes. When free, copper is toxic, mismetallating proteins and generating damaging free radicals. Consequently, copper overload is a strategy that eukaryotic cells use to combat pathogens. Bacteria have developed copper-sensing transcription factors to control copper homeostasis. The cell envelope is the first compartment that has to cope with copper stress. Dedicated two-component systems control the periplasmic response to metal overload. This paper shows that the sensor kinase of the copper-sensing two-component system present in Pseudomonadales exhibits a signal-dependent phosphatase activity controlling the activation of its cognate response regulator, distinct from previously described periplasmic Cu sensors. Importantly, the data show that the system is activated by copper levels compatible with the absence of free copper in the cell periplasm. These observations emphasize the diversity of molecular mechanisms that have evolved in bacteria to manage the copper cellular distribution.

scholarly journals Specificity of Subtilin-Mediated Activation of Histidine Kinase SpaK

2017 ◽  
Vol 83 (18) ◽  
Author(s):  
Christoph Geiger ◽  
Tobias Spieß ◽  
Sophie Marianne Korn ◽  
Peter Kötter ◽  
Karl-Dieter Entian
Keyword(s):  
Bacillus Subtilis ◽  
Histidine Kinase ◽  
Two Component System ◽  
Histidine Kinases ◽  
Component System ◽  
Content Type ◽  
Component Systems ◽  
Two Component ◽  
Specific Manner ◽  
Two Component Systems

ABSTRACT Autoinduction via two-component systems is a widespread regulatory mechanism that senses environmental and metabolic changes. Although the lantibiotics nisin and subtilin are closely related and share the same lanthionine ring structure, they autoinduce their biosynthesis in a highly specific manner. Subtilin activates only the two-component system SpaRK of Bacillus subtilis, whereas nisin activates solely the two-component system NisRK of Lactococcus lactis. To identify components that determine the specificity of subtilin autoinduction, several variants of the respective lantibiotics were analyzed for their autoinductive capacities. Here, we show that amino acid position 20 is crucial for SpaK activation, as an engineered nisin molecule with phenylalanine at position 20 (nisin N20F) was able to activate SpaK in a specific manner. In combination with the N-terminal tryptophan of subtilin (nisin I1W/N20F), SpaK autoinduction reached almost the level of subtilin-mediated autoinduction. Furthermore, the overall structure of subtilin is also important for its association with the histidine kinase. The destruction of the second lanthionine ring (subtilin C11A, ring B), as well as mutations that interfere with the flexibility of the hinge region located between lanthionine rings C and D (subtilin L21P/Q22P), abolished SpaK autoinduction. Although the C-terminal part of subtilin is needed for efficient SpaK autoinduction, the destruction of lanthionine rings D and E had no measurable impact. Based on these findings, a model for the interaction of subtilin with histidine kinase SpaK was established. IMPORTANCE Although two-component systems are important regulatory systems that sense environmental changes, very little information on the molecular mechanism of sensing or the interaction of the sensor with its respective kinase is available. The strong specificity of linear lantibiotics such as subtilin and nisin for their respective kinases provides an excellent model system to unravel the structural needs of these lantibiotics for activating histidine kinases in a specific manner. More than that, the biosyntheses of lantibiotics are autoinduced via two-component systems. Therefore, an understanding of their interactions with histidine kinases is needed for the biosynthesis of newly engineered peptide antibiotics. Using a Bacillus subtilis-based reporter system, we were able to identify the molecular constraints that are necessary for specific SpaK activation and to provide SpaK specificity to nisin with just two point mutations.

scholarly journals Osmosensing by the bacterial PhoQ/PhoP two-component system

2017 ◽  
Vol 114 (50) ◽  
pp. E10792-E10798 ◽  
Author(s):  
Jing Yuan ◽  
Fan Jin ◽  
Timo Glatter ◽  
Victor Sourjik
Keyword(s):  
Essential Role ◽  
Lateral Pressure ◽  
Low Ph ◽  
Membrane Thickness ◽  
Two Component System ◽  
Component System ◽  
Component Systems ◽  
Two Component ◽  
Two Component Systems ◽  
Tm Domain

The PhoQ/PhoP two-component system plays an essential role in the response of enterobacteria to the environment of their mammalian hosts. It is known to sense several stimuli that are potentially associated with the host, including extracellular magnesium limitation, low pH, and the presence of cationic antimicrobial peptides. Here, we show that the PhoQ/PhoP two-component systems ofEscherichia coliandSalmonellacan also perceive an osmotic upshift, another key stimulus to which bacteria become exposed within the host. In contrast to most previously established stimuli of PhoQ, the detection of osmotic upshift does not require its periplasmic sensor domain. Instead, we show that the activity of PhoQ is affected by the length of the transmembrane (TM) helix as well as by membrane lateral pressure. We therefore propose that osmosensing relies on a conformational change within the TM domain of PhoQ induced by a perturbation in cell membrane thickness and lateral pressure under hyperosmotic conditions. Furthermore, the response mediated by the PhoQ/PhoP two-component system was found to improve bacterial growth recovery under hyperosmotic stress, partly through stabilization of the sigma factor RpoS. Our findings directly link the PhoQ/PhoP two-component system to bacterial osmosensing, suggesting that this system can mediate a concerted response to most of the established host-related cues.

scholarly journals Cys303 in the Histidine Kinase PhoR Is Crucial for the Phosphotransfer Reaction in the PhoPR Two-Component System in Bacillus subtilis

2006 ◽  
Vol 189 (2) ◽  
pp. 410-421 ◽  
Author(s):  
Amr Eldakak ◽  
F. Marion Hulett
Keyword(s):  
Inorganic Phosphate ◽  
Efficient Production ◽  
Dependent Manner ◽  
Two Component System ◽  
Component System ◽  
Terminal Oxidases ◽  
Component Systems ◽  
Two Component ◽  
Two Component Systems

ABSTRACT The PhoPR two-component system activates or represses Pho regulon genes to overcome a phosphate deficiency. The Pho signal transduction network is comprised of three two-component systems, PhoPR, ResDE, and Spo0A. Activated PhoP is required for expression of ResDE from the resA promoter, while ResD is essential for 80% of Pho induction, establishing a positive feedback loop between these two-component systems to amplify the signal received by the Pho system. The role of ResD in the Pho response is via production of terminal oxidases. Reduced quinones inhibit PhoR autophosphorylation in vitro, and it was proposed that the expression of terminal oxidases leads to oxidation of the quinone pool, thereby relieving the inhibition. We show here that the reducing environment generated by dithiothreitol (DTT) in vivo inhibited Pho induction in a PhoR-dependent manner, which is in agreement with our previous in vitro data. A strain containing a PhoR variant, PhoRC303A, exhibited reduced Pho induction and remained sensitive to inhibition by DTT, suggesting that the mechanisms for Pho reduction via PhoRC303A and DTT are different. PhoR and PhoRC303A were similar with regard to cellular concentration, limited proteolysis patterns, rate of autophosphorylation, stability of PhoR∼P, and inhibition of autophosphorylation by DTT. Phosphotransfer between PhoR∼P or PhoRC303A∼P and PhoP occurred rapidly; most label from PhoR∼P was transferred to PhoP, but only 10% of the label from PhoRC303A∼P was associated with PhoP, while 90% was released as inorganic phosphate. No difference in PhoP∼P or PhoR autophosphatase activity was observed between PhoR and PhoRC303A that would explain the release of inorganic phosphate. Our data are consistent with a role for PhoRC303 in PhoR activity via stabilization of the phosphoryl-protein intermediate(s) during phosphotransfer from PhoR∼P to PhoP, which is stabilization that is required for efficient production of PhoP∼P.

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