scholarly journals Experimental Evolution Reveals Redox State Modulates Mycobacterial Pathogenicity

2021 ◽  
Author(s):  
Zheng Jiang ◽  
Zengfang Zhuang ◽  
Kaixia Mi
Keyword(s):  
Experimental Evolution ◽  
Redox State ◽  
Sigma Factors ◽  
Mutation Site ◽  
Redox States ◽  
Physiological Conditions ◽  
Component Systems ◽  
Two Component Systems ◽  
Rapid Removal ◽  
High Level

Understanding how Mycobacterium tuberculosis has evolved into a professional pathogen is helpful in studying its pathogenesis and for designing vaccines. We investigated how the evolutionary adaptation of M. smegmatis mc251 to an important clinical stressor H2O2 allows bacteria undergo coordinated genetic mutations, resulting in increased pathogenicity. Whole-genome sequencing identified a mutation site in the fur gene, which caused increased expression of katG. Using a Wayne dormancy model, mc251 showed a growth advantage over its parental strain mc2155 in recovering from dormancy under anaerobic conditions. Meanwhile, the high level of KatG in mc251 was accompanied by a low level of ATP, which meant that mc251 is at a low respiratory level. Additionally, the redox-related protein Rv1996 showed different phenotypes in different specific redox states in M. smegmatis mc2155, mc251, M. bovis BCG and M. tuberculosis mc27000. In conclusion, our study shows that the same gene presents different phenotypes under different physiological conditions. This may partly explain why M. smegmatis and M. tuberculosis have similar virulence factors and signaling transduction systems such as two-component systems and sigma factors, but due to the different redox states in the corresponding bacteria, M. smegmatis is a nonpathogen, while M. tuberculosis is a pathogen. As mc251 overcomes its shortcomings of rapid removal, it can be potentially developed as a vaccine vector.

An experimentally evolved variant of RsmA confirms its central role in the control of Pseudomonas aeruginosa social motility

2020 ◽  
Author(s):  
Sophie Robitaille ◽  
Yossef López de los Santos ◽  
Marie-Christine Groleau ◽  
Fabrice Jean-Pierre ◽  
Nicolas Doucet ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Experimental Evolution ◽  
Opportunistic Pathogen ◽  
Environmental Cues ◽  
Nucleotide Substitutions ◽  
Swarming Motility ◽  
Single Nucleotide ◽  
Component Systems ◽  
Two Component Systems

AbstractBacteria can colonize a variety of different environments by modulating their gene regulation using two-component systems. The versatile opportunistic pathogen Pseudomonas aeruginosa has been studied for its capacity to adapt to a broad range of environmental conditions. The Gac/Rsm pathway is composed of the sensor kinase GacS, that detects environmental cues, and the response regulator GacA, that modulates the expression of a specific genes. This system, through the sRNA repressors RsmY and RsmZ, negatively controls the activity of the protein RsmA, which is centrally involved in the transition from chronic to acute infections by post-transcriptionally regulating several virulence functions. RsmA positively regulates swarming motility, a social surface behaviour. Through a poorly defined mechanism, RsmA is also indirectly regulated by HptB, and a ΔhptB mutant exhibits a severe swarming defect. Since a ΔhptB mutant retains all the known functions required for that type of motility, we used an experimental evolution approach to identify elements responsible for its swarming defect. After a few passages under swarming conditions, the defect of the ΔhptB mutant was rescued by the emergence of spontaneous single nucleotide substitutions in the gacA and rsmA genes. Since GacA indirectly represses RsmA activity, it was coherent that an inactivating mutation in gacA would compensate the ΔhptB swarming defect. However, the effect of the mutation in rsmA was unexpected since RsmA promotes swarming; indeed, using expression reporters, we found that the mutation that does not abolish its activity. Instead, using electrophoretic mobility shift assays and molecular simulations, we show that this variant of RsmA is actually less amenable to titration by its cognate repressor RsmY, supporting the other phenotypes observed for this mutant. These results confirm the central role of RsmA as a regulator of swarming motility in P. aeruginosa and identify residues crucial for RsmA function in social motility.Author summaryBacteria need to readily adapt to their environment. Two-component systems (TCS) allow such adaption by triggering bacterial regulation changes through the detection of environmental cues. The opportunistic pathogen Pseudomonas aeruginosa possesses more than 60 TCS in its genome. The Gac/Rsm is a TCS extensively studied for its implication in virulence regulation. This system regulates the transition between chronic and acute bacterial infection behaviours. To acquire a better understanding of this regulation, we performed a directed experimental evolution on a swarming-deficient mutant in a poorly understood regulatory component of the Gac/Rsm pathway. We observed single nucleotide substitutions that allowed restoration of a swarming phenotype similar to the wild-type behaviour. More specifically, mutations were found in the gacA and rsmA genes. Interestingly, the observed mutation in rsmA does not result in loss of function of the protein but rather alters its susceptibility to repression by its cognate interfering sRNA. Since modification in the RNA sequence of RsmA results in the rescue of swarming motility, we confirm the central role of this posttranscriptional repressor in this social lifestyle.

scholarly journals The adaptive transcriptional response of pathogenic Leptospira to peroxide reveals new defenses against infection-related oxidative stress

2020 ◽  
Author(s):  
Crispin Zavala-Alvarado ◽  
Odile Sismeiro ◽  
Rachel Legendre ◽  
Hugo Varet ◽  
Giovanni Bussotti ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Adaptive Response ◽  
Sigma Factors ◽  
Component Systems ◽  
Two Component ◽  
Genes Encoding ◽  
Causative Agents ◽  
Two Component Systems ◽  
Non Coding Rnas ◽  
Insight Into

AbstractPathogenic Leptospira spp. are the causative agents of the waterborne zoonotic disease leptospirosis. During infection, Leptospira are confronted with dramatic adverse environmental changes such as deadly reactive oxygen species (ROS). Withstanding ROS produced by the host innate immunity is an important strategy evolved by pathogenic Leptospira for persisting in and colonizing hosts. In L. interrogans, genes encoding defenses against ROS are repressed by the peroxide stress regulator, PerR. In this study, RNA sequencing was performed to characterize both the L. interrogans adaptive response to low and high concentrations of hydrogen peroxide and the PerR regulon. We showed that Leptospira solicit three main peroxidase machineries (catalase, cytochrome C peroxidase and peroxiredoxin) and heme to detoxify oxidants produced during a peroxide stress. In addition, canonical molecular chaperones of the heat shock response and DNA repair proteins from the SOS response were required for Leptospira recovering from oxidative damages. Determining the PerR regulon allowed to identify the PerR-dependent mechanisms of the peroxide adaptive response and has revealed a PerR-independent regulatory network involving other transcriptional regulators, two-component systems and sigma factors as well as non-coding RNAs that putatively orchestrate, in concert with PerR, this adaptive response. In addition, we have identified other PerR-regulated genes encoding a TonB-dependent transport system, a lipoprotein (LipL48) and a two-component system (VicKR) involved in Leptospira tolerance to superoxide and that could represent the first defense mechanism against superoxide in L. interrogans, a bacterium lacking canonical superoxide dismutase. Our findings provide a comprehensive insight into the mechanisms required by pathogenic Leptospira to overcome infection-related oxidants during the arm race with a host. This will participate in framing future hypothesis-driven studies to identify and decipher novel virulence mechanisms in this life-threatening pathogen.Author summaryLeptospirosis is a zoonotic infectious disease responsible for over one million of severe cases and 60 000 fatalities annually worldwide. This neglected and emerging disease has a worldwide distribution, but it mostly affects populations from developing countries in sub-tropical areas. The causative agents of leptospirosis are pathogenic bacterial Leptospira spp. There is a considerable deficit in our knowledge of these atypical bacteria, including their virulence mechanisms. During infection, Leptospira are confronted with the deadly oxidants produced by the host tissues and immune response. Here, we have identified the cellular factors necessary for Leptospira to overcome the oxidative stress response. We found that Leptospira solicit peroxidases to detoxify oxidants as well as chaperones of the heat shock response and DNA repair proteins of the SOS response to recover from oxidative damage. Moreover, our study indicates that adaptation to oxidative stress is orchestrated by a regulatory network involving PerR and other transcriptional regulators, sigma factors, two component systems, and putative non-coding RNAs. These findings provide a comprehensive insight into the mechanisms required by pathogenic Leptospira to tolerate infection-related oxidants, helping identify novel virulence factors, developing new therapeutic targets and vaccines against leptospirosis.

scholarly journals A minimum set of regulators to thrive in the ocean

2020 ◽  
Vol 44 (2) ◽  
pp. 232-252
Author(s):  
S Joke Lambrecht ◽  
Claudia Steglich ◽  
Wolfgang R Hess
Keyword(s):  
Current Knowledge ◽  
Regulatory System ◽  
Sigma Factors ◽  
Marine Cyanobacteria ◽  
High Cell ◽  
Free Living ◽  
Component Systems ◽  
Two Component Systems ◽  
Ecological Success ◽  
Photosynthetic Microorganisms

ABSTRACT Marine cyanobacteria of the genus Prochlorococcus thrive in high cell numbers throughout the euphotic zones of the world's subtropical and tropical oligotrophic oceans, making them some of the most ecologically relevant photosynthetic microorganisms on Earth. The ecological success of these free-living phototrophs suggests that they are equipped with a regulatory system competent to address many different stress situations. However, Prochlorococcus genomes are compact and streamlined, with the majority encoding only five different sigma factors, five to six two-component systems and eight types of other transcriptional regulators. Here, we summarize the existing information about the functions of these protein regulators, about transcriptomic responses to defined stress conditions, and discuss the current knowledge about riboswitches, RNA-based regulation and the roles of certain metabolites as co-regulators. We focus on the best-studied isolate, Prochlorococcus MED4, but extend to other strains and ecotypes when appropriate, and we include some information gained from metagenomic and metatranscriptomic analyses.

scholarly journals Amino acid identity at one position within the α1 helix of both the histidine kinase and the response regulator of the WalRK and PhoPR two-component systems plays a crucial role in the specificity of phosphotransfer

Microbiology ◽  
2010 ◽  
Vol 156 (6) ◽  
pp. 1848-1859 ◽  
Author(s):  
Inga Jende ◽  
Kottayil I. Varughese ◽  
Kevin M. Devine
Keyword(s):  
Amino Acid ◽  
Response Regulator ◽  
Amino Acid Identity ◽  
Response Regulators ◽  
Acid Identity ◽  
Component Systems ◽  
Two Component ◽  
Two Component Systems ◽  
Interacting Surfaces ◽  
High Level

Two-component systems usually function as cognate pairs, thereby ensuring an appropriate response to the detected signal. The ability to exclusively phosphorylate a partner protein, often in the presence of many competing homologous substrates, demonstrates a high level of specificity that must derive from the interacting surfaces of the two-component system. Here, we identify positions within the histidine kinases and response regulators of the WalRK and PhoPR two-component systems of Bacillus subtilis that make a major contribution to the specificity of phosphotransfer. Changing the identity of the amino acid at position 11 within the α1 helix of WalK and at position 17 within the α1 helix of PhoP altered discrimination and allowed phosphotransfer to occur with the non-cognate partner. Changing amino acids at additional positions of the WalK kinase increased phosphotransfer, while changes at additional positions in PhoP only had an effect in the presence of the change at position 17. The importance of amino acid identity at these two positions is supported by the fact that the amino acid combinations of Ile and Ser in WalRK, and Leu and Gly in PhoPR, are very highly conserved among orthologues, while modelling indicates that these amino acid pairs are juxtaposed in the WalRK and PhoPR complexes.

scholarly journals Polymyxin Resistance ofPseudomonas aeruginosa phoQMutants Is Dependent on Additional Two-Component Regulatory Systems

2013 ◽  
Vol 57 (5) ◽  
pp. 2204-2215 ◽  
Author(s):  
Alina D. Gutu ◽  
Nicole Sgambati ◽  
Pnina Strasbourger ◽  
Mark K. Brannon ◽  
Michael A. Jacobs ◽  
...  
Keyword(s):  
Lipid A ◽  
Regulatory System ◽  
Content Type ◽  
Regulatory Systems ◽  
Clinical Resistance ◽  
Component Systems ◽  
Two Component ◽  
Two Component Systems ◽  
High Level

ABSTRACTPseudomonas aeruginosacan develop resistance to polymyxin as a consequence of mutations in the PhoPQ regulatory system, mediated by covalent lipid A modification. Transposon mutagenesis of a polymyxin-resistantphoQmutant defined 41 novel loci required for resistance, including two regulatory systems, ColRS and CprRS. Deletion of thecolRSgenes, individually or in tandem, abrogated the polymyxin resistance of a ΔphoQmutant, as did individual or tandem deletion ofcprRS. Individual deletion ofcolRorcolSin a ΔphoQmutant also suppressed 4-amino-l-arabinose addition to lipid A, consistent with the known role of this modification in polymyxin resistance. Surprisingly, tandem deletion ofcolRSorcprRSin the ΔphoQmutant or individual deletion ofcprRorcprSfailed to suppress 4-amino-l-arabinose addition to lipid A, indicating that this modification alone is not sufficient for PhoPQ-mediated polymyxin resistance inP. aeruginosa. Episomal expression ofcolRSorcprRSin tandem or ofcprRindividually complemented the Pm resistance phenotype in the ΔphoQmutant, while episomal expression ofcolR,colS, orcprSindividually did not. Highly polymyxin-resistantphoQmutants ofP. aeruginosaisolated from polymyxin-treated cystic fibrosis patients harbored mutant alleles ofcolRSandcprS; when expressed in a ΔphoQbackground, these mutant alleles enhanced polymyxin resistance. These results define ColRS and CprRS as two-component systems regulating polymyxin resistance inP. aeruginosa, indicate that addition of 4-amino-l-arabinose to lipid A is not the only PhoPQ-regulated biochemical mechanism required for resistance, and demonstrate thatcolRSandcprSmutations can contribute to high-level clinical resistance.

Diffusive layering during film growth in two-component systems with limited mutual solubility

2000 ◽  
Author(s):  
Grigory V. Merkulov ◽  
Valentin M. Ievlev ◽  
Evgeny V. Shvedov ◽  
Vadim P. Ampilogov
Keyword(s):  
Film Growth ◽  
Mutual Solubility ◽  
Component Systems ◽  
Two Component ◽  
Two Component Systems

scholarly journals Regulation of virulence and β-lactamase gene expression in Staphylococcus aureus isolates: cooperation of two-component systems in bloodstream superbugs

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Sanaz Dehbashi ◽  
Hamed Tahmasebi ◽  
Behrouz Zeyni ◽  
Mohammad Reza Arabestani
Keyword(s):  
Staphylococcus Aureus ◽  
Real Time ◽  
Real Time Pcr ◽  
Quantitative Real Time Pcr ◽  
Gene Expressions ◽  
Resistance Development ◽  
Component Systems ◽  
Two Component ◽  
Two Component Systems ◽  
Lactamase Gene

Abstract Background Methicillin-resistant Staphylococcus aureus (MRSA)-bloodstream infections (BSI) are predominantly seen in the hospital or healthcare-associated host. Nevertheless, the interactions of virulence factor (VFs) regulators and β-lactam resistance in MRSA-BSI are unclear. This study aims to characterize the molecular relationship of two-component systems of VFs and the expression of the β-lactamase gene in MRSA-BSI isolates. In this study, 639 samples were collected from BSI and identified by phenotypic methods. We performed extensive molecular characterization, including SCCmec type, agr type, VFs gene profiles determinations, and MLST on isolates. Also, a quantitative real-time PCR (q-RT PCR) assay was developed for identifying the gene expressions. Results Ninety-one (91) S. aureus and 61 MRSA (67.0%) strains were detected in BSI samples. The presence of VFs and SCCmec genes in MRSA isolates were as follows: tst (31.4%), etA (18.0%), etB (8.19%), lukS-PVL (31.4%), lukF-PV (18.0%), lukE-lukD (16.3%), edin (3.2%), hla (16.3%), hlb (18.0%), hld (14.7%), hlg (22.9%), SCCmecI (16.3%), SCCmecII (22.9%), SCCmecIII (36.0%), SCCmecIV (21.3%), and SCCmecV (16.3%). Quantitative real-time PCR showed overexpression of mecRI and mecI in the toxigenic isolates. Moreover, RNAIII and sarA genes were the highest expressions of MRSA strains. The multi-locus sequence typing data confirmed a high prevalence of CC5, CC8, and CC30. However, ST30, ST22, and ST5 were the most prevalent in the resistant and toxigenic strains. Conclusion We demonstrated that although regulation of β-lactamase gene expressions is a significant contributor to resistance development, two-component systems also influence antibiotic resistance development in MRSA-BSI isolates. This indicates that resistant strains might have pathogenic potential. We also confirmed that some MLST types are more successful colonizers with a potential for MRSA-BSI.

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.

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