scholarly journals Roles of DnaK and RpoS in Starvation-Induced Thermotolerance of Escherichia coli

1998 ◽  
Vol 180 (4) ◽  
pp. 846-854 ◽  
Author(s):  
David Rockabrand ◽  
Kevin Livers ◽  
Tess Austin ◽  
Robyn Kaiser ◽  
Debra Jensen ◽  
...  

ABSTRACT DnaK is essential for starvation-induced resistance to heat, oxidation, and reductive division in Escherichia coli. Studies reported here indicate that DnaK is also required for starvation-induced osmotolerance, catalase activity, and the production of the RpoS-controlled Dps (PexB) protein. Because thesednaK mutant phenotypes closely resemble those ofrpoS (ς38) mutants, the relationship between DnaK and RpoS was evaluated directly during growth and starvation at 30°C in strains with genetically altered DnaK content. A starvation-specific effect of DnaK on RpoS abundance was observed. During carbon starvation, DnaK deficiency reduced RpoS levels threefold, while DnaK excess increased RpoS levels nearly twofold. Complementation of the dnaK mutation restored starvation-induced RpoS levels to normal. RpoS deficiency had no effect on the cellular concentration of DnaK, revealing an epistatic relationship between DnaK and RpoS. Protein half-life studies conducted at the onset of starvation indicate that DnaK deficiency significantly destabilized RpoS. RpoH (ς32) suppressors of thednaK mutant with restored levels of RpoS and dnaK rpoS double mutants were used to show that DnaK plays both an independent and an RpoS-dependent role in starvation-induced thermotolerance. The results suggest that DnaK coordinates sigma factor levels in glucose-starved E. coli.

2021 ◽  
Author(s):  
Heesu Kim ◽  
Dong Gun Lee

Abstract Hydrogen peroxide (H2O2) is a debriding agent that damages the microbial structure and function by generating various reactive oxygen species (ROS). H2O2-produced hydroxyl radical (OH∙) also exert oxidative stress on microorganisms. The spread of antibiotic resistance in bacteria is a serious issue worldwide, and greater efforts are needed to identify and characterize novel antibacterial mechanisms to develop new treatment strategies. Therefore, this study aimed to clarify the relationship between H2O2 and Escherichia coli and to elucidate a novel antibacterial mechanism(s) of H2O2. Following H2O2 exposure, increased levels of 8-hydroxyldeoxyguanosine and malondialdehyde indicated that H2O2 accelerates oxidation of bacterial DNA and lipids in E. coli. As oxidative damage worsened, the SOS response was triggered. Cell division arrest and resulting filamentation were identified in cells, indicating that LexA was involved in DNA replication. It was also verified that RecA, a representative SOS gene, helps self-cleavage of LexA and acts as a bacterial caspase-like protein. Our findings also showed that dinF is essential to preserve E. coli from H2O2-induced ROS, and furthermore, demonstrated that H2O2-induced SOS response and SOS genes participate differently in guarding E. coli from oxidative stress. As an extreme SOS response is considered apoptosis-like death (ALD) in bacteria, additional experiments were performed to examine the characteristics of ALD. DNA fragmentation and membrane depolarization appeared in H2O2-treated cells, suggesting that H2O2 causes ALD in E. coli. In conclusion, our investigations revealed that ALD is a novel antibacterial mode of action(s) of H2O2 with important contributions from SOS genes.


2017 ◽  
Vol 80 (7) ◽  
pp. 1214-1221 ◽  
Author(s):  
Daniel L. Weller ◽  
Jasna Kovac ◽  
Sherry Roof ◽  
David J. Kent ◽  
Jeffrey I. Tokman ◽  
...  

ABSTRACT Although wildlife intrusion and untreated manure have been associated with microbial contamination of produce, relatively few studies have examined the survival of Escherichia coli on produce under field conditions following contamination (e.g., via splash from wildlife feces). This experimental study was performed to estimate the die-off rate of E. coli on preharvest lettuce following contamination with a fecal slurry. During August 2015, field-grown lettuce was inoculated via pipette with a fecal slurry that was spiked with a three-strain cocktail of rifampin-resistant nonpathogenic E. coli. Ten lettuce heads were harvested at each of 13 time points following inoculation (0, 2.5, 5, and 24 h after inoculation and every 24 h thereafter until day 10). The most probable number (MPN) of E. coli on each lettuce head was determined, and die-off rates were estimated. The relationship between sample time and the log MPN of E. coli per head was modeled using a segmented linear model. This model had a breakpoint at 106 h (95% confidence interval = 69, 142 h) after inoculation, with a daily decrease of 0.70 and 0.19 log MPN for 0 to 106 h and 106 to 240 h following inoculation, respectively. These findings are consistent with die-off rates obtained in similar studies that assessed E. coli survival on produce following irrigation. Overall, these findings provide die-off rates for E. coli on lettuce that can be used in future quantitative risk assessments.


1986 ◽  
Vol 49 (12) ◽  
pp. 944-951 ◽  
Author(s):  
J. E. KENNEDY ◽  
C. I. WEI ◽  
J. L. OBLINGER

The distribution of coliphages in various foods and the relationship between the incidences of coliphages and bacterial indicators were investigated. A total of 120 food samples comprising twelve products and including fresh meats, shellfish, vegetables and processed meats, were analyzed for indigenous coliphages using Escherichia coli hosts C, C-3000 and B. Bacterial analyses included enumeration of E. coli, fecal coliforms and coliforms, as well as aerobic plate counts and Salmonella analyses. Coliphages were detected (≥10 PFU/100 g) in 56% of samples and eleven of twelve products. Coliphages, E. coli, fecal coliforms and coliforms were recovered at a level of at least 30 organisms per 100 g in 43, 43, 68 and 81% of samples, with overall mean recoveries of 13, 19, 93 and 4300 organisms/100 g, respectively. Highest and lowest recoveries of coliphages and E. coli were from fresh meats and vacuum-packaged processed meats, respectively. Significant nonparametric correlations between coliphages, E. coli, fecal coliforms and coliforms were found among all food samples.


2020 ◽  
Vol 75 (5) ◽  
pp. 1151-1158
Author(s):  
Corey S Suelter ◽  
Nancy D Hanson

Abstract Background Virulence genes and the expression of resistance mechanisms undoubtedly play a role in the successful spread of the pandemic clone Escherichia coli ST131. Porin down-regulation is a chromosomal mechanism associated with antibiotic resistance. Translation of porin proteins can be impacted by modifications in mRNA half-life and the interaction among small RNAs (sRNAs), the porin transcript and the sRNA chaperone Hfq. Modifications in the translatability of porin proteins could impact the fitness and therefore the success of E. coli ST131 isolates in the presence of antibiotic. Objectives To identify differences in the translatability of OmpC and OmpF porins for different STs of E. coli by comparing steady-state RNA levels, mRNA half-life, regulatory sRNA expression and protein production. Methods RNA expression was evaluated using real-time RT–PCR and OmpC mRNA half-life by northern blotting. OmpC, OmpF and Hfq protein levels were evaluated by immunoblotting. Results Differences between ST131 and non-ST131 isolates included: (i) the level of OmpC RNA and protein produced with mRNA expression higher for ST131 but OmpC protein levels lower compared with non-ST131 isolates; (ii) OmpC mRNA half-life (21–30 min for ST131 isolates compared with <2–23 min for non-ST131 isolates); and (iii) levels of the sRNA MicC (2- to 120-fold for ST131 isolates compared with −4- to 70-fold for non-ST131 isolates). Conclusions Mechanisms involved in the translatability of porin proteins differed among different STs of E. coli. These differences could provide a selective advantage to ST131 E. coli when confronted with an antibiotic-rich environment.


2008 ◽  
Vol 190 (18) ◽  
pp. 6170-6177 ◽  
Author(s):  
Linda D. Rankin ◽  
Diane M. Bodenmiller ◽  
Jonathan D. Partridge ◽  
Shirley F. Nishino ◽  
Jim C. Spain ◽  
...  

ABSTRACT Chromatin immunoprecipitation and microarray (ChIP-chip) analysis showed that the nitric oxide (NO)-sensitive repressor NsrR from Escherichia coli binds in vivo to the promoters of the tynA and feaB genes. These genes encode the first two enzymes of a pathway that is required for the catabolism of phenylethylamine (PEA) and its hydroxylated derivatives tyramine and dopamine. Deletion of nsrR caused small increases in the activities of the tynA and feaB promoters in cultures grown on PEA. Overexpression of nsrR severely retarded growth on PEA and caused a marked repression of the tynA and feaB promoters. Both the growth defect and the promoter repression were reversed in the presence of a source of NO. These results are consistent with NsrR mediating repression of the tynA and feaB genes by binding (in an NO-sensitive fashion) to the sites identified by ChIP-chip. E. coli was shown to use 3-nitrotyramine as a nitrogen source for growth, conditions which partially induce the tynA and feaB promoters. Mutation of tynA (but not feaB) prevented growth on 3-nitrotyramine. Growth yields, mutant phenotypes, and analyses of culture supernatants suggested that 3-nitrotyramine is oxidized to 4-hydroxy-3-nitrophenylacetate, with growth occurring at the expense of the amino group of 3-nitrotyramine. Accordingly, enzyme assays showed that 3-nitrotyramine and its oxidation product (4-hydroxy-3-nitrophenylacetaldehyde) could be oxidized by the enzymes encoded by tynA and feaB, respectively. The results suggest that an additional physiological role of the PEA catabolic pathway is to metabolize nitroaromatic compounds that may accumulate in cells exposed to NO.


1971 ◽  
Vol 123 (4) ◽  
pp. 501-505 ◽  
Author(s):  
J. W. Dale

1. The amino acid composition of the β-lactamase from E. coli (R-1818) was determined. 2. The R-1818 β-lactamase is inhibited by formaldehyde, hydroxylamine, sodium azide, iodoacetamide, iodine and sodium chloride. 3. The Km values for benzylpenicillin, ampicillin and oxacillin have been determined by using the R-factor enzyme from different host species. The same values were obtained, irrespective of the host bacterium. 4. The molecular weight of the enzyme was found to be 44600, and was the same for all host species. 5. The relationship of R-1818 and R-GN238 β-lactamases is discussed.


2020 ◽  
Vol 86 (14) ◽  
Author(s):  
Subhrajit Bhar ◽  
Tungadri Bose ◽  
Sharmila S. Mande

ABSTRACT Signal transduction systems are essential for microorganisms to respond to their ever-changing environment. They can be distinguished into one-component systems, two-component systems, and extracytoplasmic-function σ factors. Abundances of a few signal-transducing proteins, termed herein as sensory proteins (SPs), have previously been reported to be correlated with the genome size and ecological niche of certain Gram-positive bacteria. No such reports are available for Gram-negative bacteria. The current study attempts to investigate the relationship of the abundances of SPs to genome size in Escherichia coli, and the bacterial pathotypes or phylotypes. While the relationship between SP abundance and genome size could not be established, the sensory protein index (SPI), a new metric defined herein, was found to be correlated with E. coli virulence. In addition, significant association was observed among the distribution of SPs and E. coli pathotypes. Results indicate that such associations might be due to genomic rearrangements to best utilize the resources available in a given ecological niche. Overall, the study provides an in-depth analysis of the occurrence of different SPs among pathogenic and nonpathogenic E. coli strains. Possibilities of using the SPI as a marker for identifying pathogenic strains from among an organism complex are also discussed. IMPORTANCE Sensory proteins (SPs) act as sensors and actuators for a cell and participate in important mechanisms pertaining to bacterial survival, adaptation, and virulence. Therefore, bacterial species residing in similar ecological niches or those sharing common pathotypes are expected to exhibit similar SP signatures. We have investigated profiles of SPs in different species of Escherichia coli and present in this article the sensory protein index (SPI), a metric for quantifying the abundance and/or distribution of SPs across bacterial genomes, which could indicate the virulence potency of a bacterium. The SPI could find use in characterizing uncultured strains and bacterial complexes, as a biomarker for disease diagnostics, evaluating the effect of therapeutic interventions, assessing effects of ecological alterations, etc. Grouping the studied strains of E. coli on the basis of the frequency of occurrence of SPs in their genomes could potentially replicate the stratification of these strains on the basis of their phylotypes. In addition, E. coli strains belonging to the same pathotypes were also seen to share similar SP signatures. Furthermore, the SPI was seen to be an indicator of pathogenic potency of E. coli strains. The SPI metric is expected to be useful in the (pathogenic) characterization of hereto uncultured strains which are routinely sequenced in host microbiome analysis projects, or from among an ensemble of microbial organisms constituting a biospecimen. Thus, the possibilities of using the SPI as a biomarker for diagnosis of a disease or the outcome of a therapeutic intervention cannot be ruled out. Further, SPIs obtained from longitudinal ecological samples have the potential to serve as key indicators of environmental changes. Such changes in the environment are often detrimental to the resident biome and methods for timely detection of environmental changes hold huge socioeconomic benefits.


2012 ◽  
Vol 2012 ◽  
pp. 1-10 ◽  
Author(s):  
Chin How Lee ◽  
Jack S. H. Oon ◽  
Kun Cheng Lee ◽  
Maurice H. T. Ling

Escherichia coli is commonly found in intestine of human, and any changes in their adaptation or evolution may affect the human body. The relationship between E. coli and food additives is less studied as compared to antibiotics. E. coli within our human gut are consistently interacting with the food additives; thus, it is important to investigate this relationship. In this paper, we observed the evolution of E. coli cultured in different concentration of food additives (sodium chloride, benzoic acid, and monosodium glutamate), singly or in combination, over 70 passages. Adaptability over time was estimated by generation time and cell density at stationary phase. Polymerase chain reaction (PCR)/restriction fragments length polymorphism (RFLP) using 3 primers and restriction endonucleases, each was used to characterize adaptation/evolution at genomic level. The amplification and digestion profiles were tabulated and analyzed by Nei-Li dissimilarity index. Our results demonstrate that E. coli in every treatment had adapted over 465 generations. The types of stress were discovered to be different even though different concentrations of same additives were used. However, RFLP shows a convergence of genetic distances, suggesting the presence of global stress response. In addition, monosodium glutamate may be a nutrient source and support acid resistance in E. coli.


2014 ◽  
Vol 14 (6) ◽  
pp. 1087-1094 ◽  
Author(s):  
Luis Sanchez ◽  
Lucas Guz ◽  
Pilar García ◽  
Silvia Ponce ◽  
Silvia Goyanes ◽  
...  

ZnO nanorods (ZnO NRs) were grown on ZnO seeded fluorine doped tin oxide (FTO) substrates at low temperatures (90 °C) from Zn2+ precursors in alkaline aqueous solution. The ZnO seeds were deposited on the FTO substrate heated at 350 °C by spray pyrolysis of a zinc acetate solution in a water ethanol mixture. The structure of seeds was tuned by the ethanol water ratio, Γ, which controls the solvent evaporation rate of drops impinging the substrate. The relationship between the microstructure and optical properties of the ZnO NR films and the photocatalytic antibacterial activity for Escherichia coli abatement, was determined through a detailed characterization of the material. The higher photocatalytic antibacterial activity was performed by ZnO NR films grown on seeds deposited from solutions with Γ in the 0.0–0.03 range. With these films, the population of viable E. coli dropped more than six orders, from 8 × 108 to 4 × 102 CFU. These results show the potential of these materials in water disinfection.


1983 ◽  
Vol 61 (12) ◽  
pp. 1315-1321 ◽  
Author(s):  
Peter C. Loewen ◽  
Barbara L. Triggs ◽  
Glen R. Klassen ◽  
Joel H. Weiner

A hybrid Escherichia coli: Col E1 plasmid, pLC36-19, containing a catalase gene has been identified in the Clarke and Carbon colony bank. Catalase activity was amplified two- to three-fold in the pLC36-19-containing strain relative to other hybrid-plasmid-containing strains and this activity could be induced three- or four-fold by hydrogen peroxide or ascorbic acid. The plasmid was transferred to a strain chromosomally deficient in catalase synthesis, resulting in a strain with high and inducible levels of catalase. The plasmid was also transferred to a minicell-producing strain and minicells harbouring the plasmid were found to synthesize a labelled protein with a molecular weight of 84 000 characteristic of catalase from E. coli. A catalase activity was also synthesized by the plasmid-containing minicells. Two catalase activities with associated peroxidase activities coded for by the plasmid were separable by polyacrylamide gel electrophoresis and migrated coincident with chromosomally encoded catalase–peroxidase activities. A third catalase activity which did not have an associated peroxidase activity was not coded for by the plasmid. A physical map of the 25.5-kilobase pair plasmid was constructed by restriction nuclease analysis and the relative positions of 38 restriction sites were defined.


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