Effects of sub-inhibitory concentrations of antibiotics and oxidative stress on the expression of type II toxin-antitoxin system genes in Klebsiella pneumoniae

Narimisa, Negar; Amraei, Fatemeh; Kalani, Behrooz Sadeghi; Mohammadzadeh, Rokhsareh; Jazi, Faramarz Masjedian

doi:10.1016/j.jgar.2019.09.005

Cited by 33 publications

(21 citation statements)

References 30 publications

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“…For survival in host, pathogen must overcome the oxidative stress (Forman and Torres, 2002). It is reported that multiple type II TA system genes in Klebsiella pneumoniae were involved in response to oxidative stress (Narimisa et al, 2020). In E. coli, upon oxidative stress, antitoxin MqsA is degraded and toxin MqsR was liberated, which enriched ghoT mRNAs as well as FIGURE 7 | Expression of yefMyeoB was regulated by YefM and YefM/YeoB.…”

Section: Discussionmentioning

confidence: 99%

Edwardsiella piscicida YefM-YoeB: A Type II Toxin-Antitoxin System That Is Related to Antibiotic Resistance, Biofilm Formation, Serum Survival, and Host Infection

DongMei

Shi

et al. 2021

Front. Microbiol.

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The emergence of drug resistant bacteria is a tricky and confronted problem in modern medicine, and one of important reasons is the widespread of toxin-antitoxin (TA) systems in pathogenic bacteria. Edwardsiella piscicida (also known as E. tarda) is the leading pathogen threatening worldwide fresh and seawater aquaculture industries and has been considered as a model organism for studying intracellular and systemic infections. However, the role of type II TA systems are completely unknown in aquatic pathogenic bacteria. In this study, we identified and characterized a type II TA system, YefM-YoeB, of E. piscicida, where YefM is the antitoxin and YoeB is the toxin. yefM and yoeB are co-expressed in a bicistronic operon. When expressed in E. coli, YoeB cause bacterial growth arrest, which was restored by the addition of YefM. To investigate the biological role of the TA system, two markerless yoeB and yefM-yoeB in-frame mutant strains, TX01ΔyoeB and TX01ΔyefM-yoeB, were constructed, respectively. Compared to the wild strain TX01, TX01ΔyefM-yoeB exhibited markedly reduced resistance against oxidative stress and antibiotic, and markedly reduced ability to form persistent bacteria. The deletion of yefM-yoeB enhanced the bacterial ability of high temperature tolerance, biofilm formation, and host serum resistance, which is the first study about the relationship between type II TA system and serum resistance. In vitro infection experiment showed that the inactivation of yefM-yoeB greatly enhanced bacterial capability of adhesion in host cells. Consistently, in vivo experiment suggested that the yefM-yoeB mutation had an obvious positive effect on bacteria dissemination of fish tissues and general virulence. Introduction of a trans-expressed yefM-yoeB restored the virulence of TX01ΔyefM-yoeB. These findings suggest that YefM-YoeB is involved in responding adverse circumstance and pathogenicity of E. piscicida. In addition, we found that YefM-YoeB negatively autoregulated the expression of yefM-yoeB and YefM could directly bind with own promoter. This study provides first insights into the biological activity of type II TA system YefM-YoeB in aquatic pathogenic bacteria and contributes to understand the pathogenesis of E. piscicida.

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Section: Discussionmentioning

confidence: 99%

Edwardsiella piscicida YefM-YoeB: A Type II Toxin-Antitoxin System That Is Related to Antibiotic Resistance, Biofilm Formation, Serum Survival, and Host Infection

DongMei

Shi

et al. 2021

Front. Microbiol.

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“…It was proposed that the fitness advantage of most bacteria is due to the costs associated with expression of TA genes. TA systems are abundantly existing in bacterial genomes and under stress conditions, toxin can target diverse cellular processes, such as translation, transcription, replication, and cell wall synthesis [8,21]. This process leading to the inhibition of cell growth, and switch to a dormant state, which cause to responding to stress conditions [22,23].…”

Section: In the Study Of Chan Et Al (2018) Indicate Yefm-yoeb And Relmentioning

confidence: 99%

“…A total volume of 20 μl reaction containing 2 μl of cDNA, 12.5 μl SYBR Green master mix, 3.5 μl nuclease-free water, and 1 μl of each primer (5 pmol) was performed according to the following program: an initial activation step at 94 °C for 10min, 45 cycles of denaturation at 95 °C for 30 s, and one cycle of 60 ℃ for 45 s. the 16s rRNA was used as an internal control to normalization of mRNA levels and fold changes. Expression was calculated by the 2 -Δ ΔCT method [8,15].…”

Section: Rna Isolation and Quantitative Real-time Reverse-transcriptamentioning

confidence: 99%

“…Unfavorable conditions such as starvation, antibiotic stresses and heat shock, resulting in degradation of unstable antitoxin, and the imbalance between toxin and antitoxin, lead to toxin interferes with vital cellular processes such as replication, transcription, translation and cell wall synthesis [4,7]. Therefore, cellular growth decreases and induces diverse responses to stress conditions [8].…”

Section: Introductionmentioning

confidence: 99%

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Expression of Type II Toxin/Antitoxin systems and ClpP protease of Methicillin-Resistant Staphylococcus aureus under stress condition

Karimaei

Shahrokhi

et al. 2020

Preprint

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BackgroundStaphylococcus aureus is a major human pathogen causing chronic to persistent infections. Amongst diverse factors of pathogenesis in bacteria, toxin-antitoxin (TA) systems have a potential to be presented as an antibacterial target due to their participation in cell physiology including stress responses. This study was conducted to determine the effects of thermal and oxidative stresses on expression of type II Toxin/Antitoxin systems and ClpP protease in Methicillin-Resistant Staphylococcus aureus (MRSA).Materials/methodsExpression of type II TA genes (mazF, relE1, relE2 and immA) and clpP gene in MRSA strain were evaluated following thermal and oxidative stresses by qRT-PCR techniques.ResultsThe cell viability was constant across thermal stress, whereas oxidative stress induction resulted in a significant reduction in the growth of MRSA strain. The result of RT-qPCR revealed that TA genes were expressed in stress conditions and expression of mazF gene increased under both thermal and oxidative stresses in MRSA strain.ConclusionsBased on our results, the MRSA strain responded to stress by altering the expression level of TA genes. In overall, TA system could be an antibacterial target in S. aureus that can revitalize the research on TA systems in this pathogen.

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“…Several mechanisms contribute to the formation of persister cells, one of which is the TA systems, and in particular type II TA systems. Toxin and antitoxin of type II TA systems are both proteins with a wide distribution in bacteria [4][5][6]. The mechanism of TA systems is based on a stable toxin and a labile antitoxin located on a plasmid or chromosome.…”

Section: Introductionmentioning

confidence: 99%

The Expression of Type II TA System Genes Following Persister Cell Formation in Pseudomonas Aeruginosa Isolates in The Exponential and Stationary Phases

Zadeh

Mirshekar

Kalani

et al. 2021

Preprint

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Objectives: Failure of infection therapy in the presence of antibiotics has become a major problem which has been mostly attributed to the ability of bacterial persister cell formation. Bacteria use various mechanisms to form persister cells in different phases, among which is the toxin-antitoxin (TA) systems. This study aimed at investigating the expression of type II TA system genes under the stress of ciprofloxacin and colistin antibiotics in the exponential and stationary phases.Methods: To determine the effects of ciprofloxacin and colistin on persister cell formation in the exponential and stationary phases of Pseudomonas aeruginosa strains, colony counting was performed at different time intervals in the presence of 5-fold MIC of ciprofloxacin and colistin. In addition, the expression of relBE, Xre-COG5654, vapBC, and Xre-GNAT genes in P. aeruginosa isolates was assessed 3.5 h after antibiotic treatment in the exponential and stationary phases using qRT-PCR.Results: Our results indicated the presence of persister phenotype of P. aeruginosa strains in the presence of 5-fold MIC of ciprofloxacin and colistin compared to the control after 3.5 h of incubation in the exponential and stationary phases. Also, the number of persister cells in the stationary phase was higher than that of the exponential phase. According to the results of qRT-PCR, ciprofloxacin and colistin may induce persister cells by increasing the expression of type II TA systems in stationary and exponential phases.Conclusion: Ciprofloxacin and colistin may increase the formation of persister cells by affecting the expression of type II TA systems.

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Effects of sub-inhibitory concentrations of antibiotics and oxidative stress on the expression of type II toxin-antitoxin system genes in Klebsiella pneumoniae

Cited by 33 publications

References 30 publications

Edwardsiella piscicida YefM-YoeB: A Type II Toxin-Antitoxin System That Is Related to Antibiotic Resistance, Biofilm Formation, Serum Survival, and Host Infection

Edwardsiella piscicida YefM-YoeB: A Type II Toxin-Antitoxin System That Is Related to Antibiotic Resistance, Biofilm Formation, Serum Survival, and Host Infection

Expression of Type II Toxin/Antitoxin systems and ClpP protease of Methicillin-Resistant Staphylococcus aureus under stress condition

The Expression of Type II TA System Genes Following Persister Cell Formation in Pseudomonas Aeruginosa Isolates in The Exponential and Stationary Phases

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