scholarly journals A Universal Stress Protein upregulated by hypoxia may contribute to chronic lung colonisation and intramacrophage survival in cystic fibrosis

2020 ◽  
Author(s):  
Andrew O’Connor ◽  
Rita Berisio ◽  
Mary Lucey ◽  
Kirsten Schaffer ◽  
Siobhán McClean
Keyword(s):  
Oxidative Stress ◽  
Chronic Infection ◽  
Stress Proteins ◽  
Cell Attachment ◽  
Stress Protein ◽  
Acid Stress ◽  
Nutrient Deficiency ◽  
Fold Increase ◽  
Burkholderia Cenocepacia ◽  
Low Oxygen

SummaryUniversal stress proteins (USPs) are ubiquitously expressed in bacteria, plants and eukaryotes and play a lead role in adaptation to environmental conditions. In Gram negative bacteria they enable adaption of bacterial pathogens to the conditions encountered in the human niche, including hypoxia, oxidative stress, osmotic stress, nutrient deficiency or acid stress, thereby facilitating colonisation. We previously reported that all six USP proteins encoded within a low-oxygen responsive locus in Burkholderia cenocepacia showed increased abundance during chronic colonisation of the CF lung. However, the role of USPs in chronic infection is not known. Using mutants derived from B. cenocepacia strain, K56-2, we show that USP76 is required for growth and survival in many conditions associated with the CF lung including, hypoxia, acidic conditions, oxidative stress. Moreover, it is involved in attachment to host epithelial cells, but not virulence. It also has a role in survival in macrophages isolated from people with CF. In contrast, another USP encoded in the same locus, USP92 had no effect on host cell attachment or oxidative stress, but was responsible for a 3-fold increase in virulence. Overall this shows that these USPs, both upregulated during chronic infection, have distinct roles in Burkholderia pathogenesis and may support the survival of B. cenocepacia in the CF lung. Specifically, USP76 is involved in its survival within CF macrophages, a hallmark of Burkholderia infection.

Induction of HSP 32 gene in hypoxic cardiomyocytes is attenuated by treatment withN-acetyl-l-cysteine

1998 ◽  
Vol 274 (3) ◽  
pp. H965-H973 ◽  
Author(s):  
D. R. Borger ◽  
D. A. Essig
Keyword(s):  
Heme Oxygenase ◽  
Stress Proteins ◽  
Stress Protein ◽  
Heme Oxygenase 1 ◽  
Low Oxygen ◽  
Neonatal Cardiomyocytes ◽  
Ambient Oxygen ◽  
Mrna Content ◽  
Mrna And Protein Expression ◽  
Heat Stress Protein

Increased synthesis of stress proteins may enhance myocardial viability during periods of low oxygen delivery. Our purpose was to determine if the oxidative stress protein heme oxygenase-1 [heat stress protein 32 (HSP 32)] was induced in hypoxic cardiomyocytes and whether this induction might be mediated by a redox-sensitive mechanism. Primary rat neonatal cardiomyocytes, cultured to express a tissuelike phenotype, responded to 12 h of hypoxia (<0.5% ambient oxygen) with an approximately fivefold (range 3- to 7.5-fold; P < 0.05) increase in HSP 32 mRNA and a threefold ( P < 0.05) increase in HSP 32 protein content. Exposure to 80 μM H2O2for 3 h increased HSP 32 mRNA content to a similar extent. Expression of heme oxygenase-2 mRNA was unaffected by H2O2or hypoxic treatments. Inclusion of 20 mM N-acetyl-l-cysteine in the medium during hypoxia reduced the increase in HSP 32 mRNA and protein expression by 25–50% compared with hypoxia alone. The data suggest that induction of HSP 32 protein may lead to an improved antioxidant defense in cardiomyocytes during hypoxia and that a redox-sensitive pathway mediates at least a portion of the hypoxic induction of the HSP 32gene.

Induction of heme oxygenase-1 (HSP32) mRNA in skeletal muscle following contractions

1997 ◽  
Vol 272 (1) ◽  
pp. C59-C67 ◽  
Author(s):  
D. A. Essig ◽  
D. R. Borger ◽  
D. A. Jackson
Keyword(s):  
Oxidative Stress ◽  
Stress Proteins ◽  
Stress Protein ◽  
Polymerase Chain Reaction Analysis ◽  
Relative Increase ◽  
Muscle Contractions ◽  
Heme Oxygenase 1 ◽  
Control Value ◽  
Actin Mrna ◽  
Heat Stress Proteins

The capacity of preexisting antioxidant pathways to handle oxidative stress during exercise may be complemented by the synthesis of inducible heat stress proteins (HSP). Our purpose was to determine if the amount of mRNA for HSP32, a major oxidative stress protein, was increased in muscle after repetitive contractions. Reverse transcriptase-polymerase chain reaction analysis showed that HSP32 mRNA (normalized to alpha-actin mRNA) was increased about seven- and about fourfold (P < 0.35) immediately after 1 h of exhaustive running and after 3 h of muscle contractions (10 Hz nerve stimulation), respectively. Northern blot analysis revealed that HSP70 mRNAs were 3.5- to 15.5-fold above control value (P < 0.05), whereas the content of another oxidative stress protein mRNA (macrophage stress protein 23) was unchanged 0 h after contractions. The relative increase in HSP32 mRNA was found to be dependent on active tension generation; passive tension did not increase the HSP32-to-actin mRNA ratio. Increases in HSP32 mRNA may underlie an inducible antioxidant pathway in muscle responsive to metabolic stresses associated with repeated muscle contractions.

scholarly journals The DNA mimic protein BCAS0292 is involved in the regulation of virulence of Burkholderia cenocepacia

2020 ◽  
Author(s):  
Ruth Dennehy ◽  
Simon Dignam ◽  
Sarah McCormack ◽  
Maria Romano ◽  
Yueran Hou ◽  
...  
Keyword(s):  
Cell Attachment ◽  
Regulation Of Gene Expression ◽  
Dna Supercoiling ◽  
Burkholderia Cenocepacia ◽  
Opportunistic Pathogens ◽  
Global Regulator ◽  
Low Oxygen ◽  
Growth And Survival ◽  
Hypoxic Environment ◽  
Host Cell Attachment

AbstractAdaptation of opportunistic pathogens to their host environment requires reprogramming of a vast array of genes to facilitate survival in the host. Burkholderia cenocepacia, a Gram-negative bacterium that colonizes environmental niches, is exquisitely adaptable to the hypoxic environment of the cystic fibrosis lung and survives in macrophages. B. cenocepacia possesses a large genome encoding multiple virulence systems, stress response proteins and a large locus that responds to low oxygen. We previously identified BCAS0292, an acidic protein encoded on replicon 3. Deletion of the BCAS0292 gene resulted in altered abundance of >1000 proteins; 46 proteins became undetectable while 556 proteins showed ≥1.5-fold reduced abundance, suggesting BCAS0292 is a global regulator. Moreover, the ΔBCAS0292 mutant showed a range of pleiotropic effects: virulence, host-cell attachment and motility were reduced, antibiotic susceptibility was altered and biofilm formation enhanced. Its growth and survival were impaired in 6% oxygen. Structural analysis revealed BCAS0292 presents a dimeric β-structure with a negative electrostatic surface. Further, the ΔBCAS0292 mutant displayed altered DNA supercoiling, implicated in global regulation of gene expression. We propose that BCAS0292 acts as a DNA-mimic, altering DNA topology and regulating the expression of multiple genes, thereby enabling the adaptation of B. cenocepacia to highly diverse environments.

scholarly journals Differential Roles of the Universal Stress Proteins of Escherichia coli in Oxidative Stress Resistance, Adhesion, and Motility

2005 ◽  
Vol 187 (18) ◽  
pp. 6265-6272 ◽  
Author(s):  
Laurence Nachin ◽  
Ulf Nannmark ◽  
Thomas Nyström
Keyword(s):  
Oxidative Stress ◽  
Escherichia Coli ◽  
Stress Resistance ◽  
Stress Proteins ◽  
Stress Protein ◽  
Physiological Role ◽  
Cellular Growth ◽  
Yeast Cells ◽  
Phenotypic Characterization ◽  
Oxidative Stress Resistance

ABSTRACT The universal stress protein (UspA) superfamily encompasses a conserved group of proteins that are found in bacteria, archaea, and eukaryotes. Escherichia coli harbors six usp genes—uspA, -C, -D, -E, -F, and -G—the expression of which is triggered by a large variety of environmental insults. The uspA gene is important for survival during cellular growth arrest, but the exact physiological role of the Usp proteins is not known. In this work we have performed phenotypic characterization of mutants with deletions of the six different usp genes. We report on hitherto unknown functions of these genes linked to motility, adhesion, and oxidative stress resistance, and we show that usp functions are both overlapping and distinct. Both UspA and UspD are required in the defense against superoxide-generating agents, and UspD appears also important in controlling intracellular levels of iron. In contrast, UspC is not involved in stress resistance or iron metabolism but is essential, like UspE, for cellular motility. Electron microscopy demonstrates that uspC and uspE mutants are devoid of flagella. In addition, the function of the uspC and uspE genes is linked to cell adhesion, measured as FimH-mediated agglutination of yeast cells. While the UspC and UspE proteins promote motility at the expense of adhesion, the UspF and UspG proteins exhibit the exact opposite effects. We suggest that the Usp proteins have evolved different physiological functions that reprogram the cell towards defense and escape during cellular stress.

Increased oxidative stress and plasma Hsp70 levels among gasoline filling station attendants

2016 ◽  
Vol 33 (2) ◽  
pp. 171-181 ◽  
Author(s):  
Bing Xia ◽  
Kangcheng Chen ◽  
Yingnan Lv ◽  
Damin Huang ◽  
Jing Liu ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Stress Protein ◽  
Weighted Average ◽  
Exposed Group ◽  
Cumulative Exposure ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Gasoline Station ◽  
Filling Station ◽  
Exposed Workers

Objectives: Methylcyclopentadienyl manganese tricarbonyl (MMT) is an organic derivative of manganese (Mn) and is used as an antiknock agent and octane enhancer in gasoline. In this article, we tested the oxidative stress and heat stress protein (Hsp) 70 levels of gasoline station attendants to explore potential plasma biomarkers. Furthermore, the dose–response relationship was also identified. Methods: A total of 144 workers, including 96 petrol fillers and 48 cashiers, participated in the study. Ambient concentrations of benzene, toluene, ethylbenzene, and xylene (BTEX) and Mn were monitored at nine filling stations. During the measuring process, the individual cumulative exposure index was calculated. Plasma oxidative stress and Hsp70 levels were also analysed using enzyme-linked immunosorbent assay. Results: The BTEX time-weighted average in office areas was significantly lower than in refuelling areas ( p < 0.05). In refuelling areas, the content of Mn ranged from 6.44 μg/m3 to 127.34 μg/m3, which was much higher than that in office areas (3.16–7.22 μg/m3; p < 0.05). Exposed workers had significantly different plasma oxidative stress indicators compared with the control group, respectively: superoxide dismutase (SOD), 39.18 ± 6.05 U/mL versus 52.84 ± 3.87 U/mL; glutathione peroxidase (GSH-Px), 186.07 ± 15.63 U versus 194.38 ± 10.42 U; and malondialdehyde (MDA), 1.68 ± 0.52 nmol/L versus 1.43 ± 0.64 nmol/L (in all comparisons, p < 0.05). Plasma Hsp70 level in the exposed group (2.77 ± 0.64 ng/mL) was significantly higher than in the control group (2.32 ± 0.87 ng/mL; p < 0.05). Furthermore, Hsp70 levels were inversely correlated with the activities of SOD ( r = −0.305) and GSH-Px ( r = −0.302) in the exposed group ( p < 0.05). Moreover, a positive correlation ( r = 0.653) was found between plasma Hsp70 levels and plasma MDA levels ( p < 0.05). Conclusion: Exposure to MMT-containing gasoline may result in increasing reactive oxygen stress among filling station attendants. Plasma Hsp70 levels could be used as a sensitive responsive biomarker for exposed workers.

MP66-05 OXIDATIVE STRESS PROTEINS IDENTIFIED IN HUMAN SPERMATOZOA BY PROTEOMIC AND BIOINFORMATIC ANALYSIS

2014 ◽  
Vol 191 (4S) ◽  
Author(s):  
Rakesh Sharma ◽  
Ashok Agarwal ◽  
Damayanthi Durairajanayagam ◽  
Zhihong Cui ◽  
Ahmet Ayaz ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Stress Proteins ◽  
Bioinformatic Analysis ◽  
Human Spermatozoa

scholarly journals Characterization of the thermotolerant cell. I. Effects on protein synthesis activity and the regulation of heat-shock protein 70 expression.

1988 ◽  
Vol 106 (4) ◽  
pp. 1105-1116 ◽  
Author(s):  
L A Mizzen ◽  
W J Welch
Keyword(s):  
Protein Synthesis ◽  
Heat Shock ◽  
Stress Proteins ◽  
Stress Protein ◽  
Shock Treatment ◽  
Heat Shock Treatment ◽  
Normal Medium ◽  
Growth Environment ◽  
Mammalian Cell Lines ◽  
C 30

Exposure of mammalian cells to a nonlethal heat-shock treatment, followed by a recovery period at 37 degrees C, results in increased cell survival after a subsequent and otherwise lethal heat-shock treatment. Here we characterize this phenomenon, termed acquired thermotolerance, at the level of translation. In a number of different mammalian cell lines given a severe 45 degrees C/30-min shock and then returned to 37 degrees C, protein synthesis was completely inhibited for as long as 5 h. Upon resumption of translational activity, there was a marked induction of heat-shock (or stress) protein synthesis, which continued for several hours. In contrast, cells first made thermotolerant (by a pretreatment consisting of a 43 degrees C/1.5-h shock and further recovery at 37 degrees C) and then presented with the 45 degrees C/30-min shock exhibited considerably less translational inhibition and an overall reduction in the amount of subsequent stress protein synthesis. The acquisition and duration of such "translational tolerance" was correlated with the expression, accumulation, and relative half-lives of the major stress proteins of 72 and 73 kD. Other agents that induce the synthesis of the stress proteins, such as sodium arsenite, similarly resulted in the acquisition of translational tolerance. The probable role of the stress proteins in the acquisition of translational tolerance was further indicated by the inability of the amino acid analogue, L-azetidine 2-carboxylic acid, an inducer of nonfunctional stress proteins, to render cells translationally tolerant. If, however, analogue-treated cells were allowed to recover in normal medium, and hence produce functional stress proteins, full translational tolerance was observed. Finally, we present data indicating that the 72- and 73-kD stress proteins, in contrast to the other major stress proteins (of 110, 90, and 28 kD), are subject to strict regulation in the stressed cell. Quantitation of 72- and 73-kD synthesis after heat-shock treatment under a number of conditions revealed that "titration" of 72/73-kD synthesis in response to stress may represent a mechanism by which the cell monitors its local growth environment.

scholarly journals Effects of the Bacterial Extract OM-85 on Phagocyte Functions and the Stress Response

1994 ◽  
Vol 3 (2) ◽  
pp. 143-148 ◽  
Author(s):  
S. Baladi ◽  
S. Kantengwa ◽  
Y. R. A. Donati ◽  
B. S. Polla
Keyword(s):  
Stress Response ◽  
Respiratory Burst ◽  
Stress Proteins ◽  
Human Peripheral Blood ◽  
Stress Protein ◽  
Blood Monocytes ◽  
Bacterial Extract ◽  
Heat Shock Stress ◽  
Intracellular Expression ◽  
Glucose Regulated Protein

The effects of the bacterial extract OM-85 on the respiratory burst, intracellular calcium and the stress response have been investigated in human peripheral blood monocytes from normal donors. Activation of the respiratory burst during bacterial phagocytosis has been previously associated with heat shock/stress proteins synthesis. Whereas OM-85 stimulated superoxide production and increased Ca2+mobilization, it fared to induce synthesis of classical HSPs. The lack of stress protein induction was observed even in the presence of iron which potentiates both oxidative injury and stress protein induction during bacterial phagocytosis. However OM-85 induced a 75–78 kDa protein, which is likely to be a glucose regulated protein (GRP78), and enhanced intracellular expression of interleukin-lβ precursor.

scholarly journals Responses of hypertrophied myocytes to reactive species: implications for glycolysis and electrophile metabolism

2011 ◽  
Vol 435 (2) ◽  
pp. 519-528 ◽  
Author(s):  
Brian E. Sansbury ◽  
Daniel W. Riggs ◽  
Robert E. Brainard ◽  
Joshua K. Salabei ◽  
Steven P. Jones ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Oxygen Consumption ◽  
Energy Demand ◽  
Lactate Production ◽  
Fold Increase ◽  
Cellular Protein ◽  
Reactive Species ◽  
Neonatal Rat ◽  
Glycolytic Flux ◽  
Rat Cardiomyocytes

During cardiac remodelling, the heart generates higher levels of reactive species; yet an intermediate ‘compensatory’ stage of hypertrophy is associated with a greater ability to withstand oxidative stress. The mechanisms underlying this protected myocardial phenotype are poorly understood. We examined how a cellular model of hypertrophy deals with electrophilic insults, such as would occur upon ischaemia or in the failing heart. For this, we measured energetics in control and PE (phenylephrine)-treated NRCMs (neonatal rat cardiomyocytes) under basal conditions and when stressed with HNE (4-hydroxynonenal). PE treatment caused hypertrophy as indicated by augmented atrial natriuretic peptide and increased cellular protein content. Hypertrophied myocytes demonstrated a 2.5-fold increase in ATP-linked oxygen consumption and a robust augmentation of oligomycin-stimulated glycolytic flux and lactate production. Hypertrophied myocytes displayed a protected phenotype that was resistant to HNE-induced cell death and a unique bioenergetic response characterized by a delayed and abrogated rate of oxygen consumption and a 2-fold increase in glycolysis upon HNE exposure. This augmentation of glycolytic flux was not due to increased glucose uptake, suggesting that electrophile stress results in utilization of intracellular glycogen stores to support the increased energy demand. Hypertrophied myocytes also had an increased propensity to oxidize HNE to 4-hydroxynonenoic acid and sustained less protein damage due to acute HNE insults. Inhibition of aldehyde dehydrogenase resulted in bioenergetic collapse when myocytes were challenged with HNE. The integration of electrophile metabolism with glycolytic and mitochondrial energy production appears to be important for maintaining myocyte homoeostasis under conditions of increased oxidative stress.

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